[发明专利]嵌合蛋白pAgoE及构建方法、应用以及使用向导的嵌合蛋白pAgoE及构建方法、应用

权利要求书

1.一种嵌合蛋白pAgoE，其特征在于：所述的嵌合蛋白pAgoE含有具有基因组靶向定位功能的pAgo结构域，以及与所述pAgo结构域连接的、具有基因组切割或修饰活性的效应结构域E；

所述pAgo结构域与任意一种使用向导的天然原核Argonaute蛋白的氨基酸序列同源性大于30％，且：

所述pAgo结构域采用来自Rhodobacter sphaeroides ATCC 17025的、NCBI序列号为ABP72561.1的RsAgo，以构建得到RsAgoE型的嵌合蛋白pAgoE；

或者，所述pAgo结构域采用来自Marinitoga piezophila的、NCBI序列号为WP_014295921.1的MpAgo，以构建得到MpAgoE型的嵌合蛋白pAgoE；

或者，所述pAgo结构域采用MpAgo的PIWI结构域核酸酶活性失活的人工突变体dMpAgo，以构建得到dMpAgoE型的嵌合蛋白pAgoE；

或者，所述pAgo结构域采用NgAgo的PIWI结构域核酸酶失活的人工突变体dNgAgo，以构建得到的dNgAgoE型的嵌合蛋白pAgoE；

其中，构建得到的RsAgoE型的嵌合蛋白pAgoE的氨基酸序列为：

Met Ala Pro Lys Lys Lys Arg Lys Val Gly Ile His Gly Val Pro Ala Ala SerSer Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys LeuLys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser Thr GlnAsp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met Lys Val Tyr Gly Tyr ArgGly Lys His Leu Gly Gly Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly SerPro Ile Asp Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn LeuPro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg AsnLys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr Glu PheLys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys Ala Gln Leu Thr ArgLeu Asn His Ile Thr Asn Cys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu IleGly Gly Glu Met Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys PheAsn Asn Gly Glu Ile Asn Phe Gly Gly Gly Gly Ser Met Ala Pro Val Gln Ala AlaAsp Glu Met Tyr Asp Ser Asn Pro His Pro Asp Arg Arg Gln Leu Val Ser Asn GlyPhe Glu Val Asn Leu Pro Asp Gln Val Glu Val Ile Val Arg Asp Leu Pro Asp ProSer Lys Val Lys Glu Glu Arg Thr Arg Leu Met Gly Tyr Trp Phe Val His Trp PheAsp Gly Lys Leu Phe His Leu Arg Ile Lys Ala Gly Gly Pro Asn Val Asp Gly GluHis Arg Ala Ile Arg Thr Ala Glu His Pro Trp Leu Leu Arg Ala Arg Leu Asp AspAla Leu Glu Glu Ala Leu Pro Lys Tyr Ala Ala Val Lys Lys Arg Pro Phe Thr PheLeu Ala Gln Lys Asp Glu Leu Ile Asp Ala Ala Ala Thr Ala Ala Gly Leu Ser HisArg Leu Leu Asn Ser Phe Lys Val Ile Pro Arg Phe Ala Leu Ser Pro Lys Ile TyrGlu Pro Val Asp Gly Thr Thr Arg Val Gly Val Phe Val Thr Ile Gly Met Arg TyrAsp Ile Glu Ala Ser Leu Arg Asp Leu Leu Glu Ala Gly Ile Asp Leu Arg Gly MetTyr Val Val Arg Arg Lys Arg Gln Pro Gly Glu Arg Gly Leu Leu Gly Arg Val ArgAla Ile Ser Asp Asp Met Val Gln Leu Phe Glu Glu Thr Asp Leu Ala Ser Val AsnVal Asn Asp Ala Lys Leu Glu Gly Ser Lys Glu Asn Phe Thr Arg Cys Leu Ser AlaLeu Leu Gly His Asn Tyr Lys Lys Leu Leu Asn Ala Leu Asp Asp Gln Glu Ala GlyTyr Arg Thr Gly Pro Arg Phe Asp Asp Ala Val Arg Arg Met Gly Glu Phe Leu AlaLys Lys Pro Ile Arg Leu Ala Asp Asn Ile Asn Ala Gln Val Gly Asp Arg Ile ValPhe Ser Asn Glu Gly Gln Ala Arg Asn Val Arg Leu Ala Pro Lys Val Glu Tyr ValPhe Asp Arg Thr Gly Ala Lys Ser Ala Glu Tyr Ala Trp Arg Gly Leu Ser Gln PheGly Pro Phe Asp Arg Pro Ser Phe Ala Asn Arg Ser Pro Arg Ile Leu Val Val TyrPro Ser Ser Thr Gln Gly Lys Val Glu Asn Phe Leu Ser Ala Phe Arg Asp Gly MetGly Ser Asn Tyr Ser Gly Phe Ser Lys Gly Phe Val Asp Leu Met Gly Leu Thr LysVal Glu Phe Val Met Cys Pro Val Glu Val Ser Ser Ala Asp Arg Asn Gly Ala HisThr Lys Tyr Asn Ser Ala Ile Glu Asp Lys Leu Ala Gly Ala Gly Glu Val His AlaGly Ile Val Val Leu Phe Glu Asp His Ala Arg Leu Pro Asp Asp Arg Asn Pro TyrIle His Thr Lys Ser Leu Leu Leu Thr Leu Gly Val Pro Thr Gln Gln Val Arg MetPro Thr Val Leu Leu Glu Pro Lys Ser Leu Gln Tyr Thr Leu Gln Asn Phe Ser IleAla Thr Tyr Ala Lys Leu Asn Gly Thr Pro Trp Thr Val Asn His Asp Lys Ala IleAsn Asp Glu Leu Val Val Gly Met Gly Leu Ala Glu Leu Ser Gly Ser Arg Thr GluLys Arg Gln Arg Phe Val Gly Ile Thr Thr Val Phe Ala Gly Asp Gly Ser Tyr LeuLeu Gly Asn Val Ser Lys Glu Cys Glu Tyr Glu Gly Tyr Ser Asp Ala Ile Arg GluSer Met Thr Gly Ile Leu Arg Glu Leu Lys Lys Arg Asn Asn Trp Arg Pro Gly AspThr Val Arg Val Val Phe His Ala His Arg Pro Leu Lys Arg Val Asp Val Ala SerIle Val Phe Glu Cys Thr Arg Glu Ile Gly Ser Asp Gln Asn Ile Gln Met Ala PheVal Thr Val Ser His Asp His Pro Phe Val Leu Ile Asp Arg Ser Glu Arg Gly LeuGlu Ala Tyr Lys Gly Ser Thr Ala Arg Lys Gly Val Phe Ala Pro Pro Arg Gly AlaIle Ser Arg Val Gly Arg Leu Thr Arg Leu Leu Ala Val Asn Ser Pro Gln Leu IleLys Arg Ala Asn Thr Pro Leu Pro Thr Pro Leu Leu Val Ser Leu His Pro Asp SerThr Phe Lys Asp Val Asp Tyr Leu Ala Glu Gln Ala Leu Lys Phe Thr Ser Leu SerTrp Arg Ser Thr Leu Pro Ala Ala Thr Pro Val Thr Ile Phe Tyr Ser Glu Arg IleAla Glu Leu Leu Gly Arg Leu Lys Ser Ile Pro Asn Trp Ser Ser Ala Asn Leu AsnIle Lys Leu Lys Trp Ser Arg Trp Phe Leu；

或者，构建得到的RsAgoE型的嵌合蛋白pAgoE的氨基酸序列为：Met Ala Pro Lys LysLys Arg Lys Val Gly Ile His Gly Val Pro Ala Ala Ser Ser Gln Leu Val Lys SerGlu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys Tyr Val Pro His GluTyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu MetLys Val Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly GlySer Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly ValIle Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln Ala AspGlu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His Ile Asn Pro AsnGlu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val SerGly His Phe Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr AsnCys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile LysAla Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile AsnPhe Gly Gly Gly Gly Ser Met Ala Pro Val Gln Ala Ala Asp Glu Met Tyr Asp SerAsn Pro His Pro Asp Arg Arg Gln Leu Val Ser Asn Gly Phe Glu Val Asn Leu ProAsp Gln Val Glu Val Ile Val Arg Asp Leu Pro Asp Pro Ser Lys Val Lys Glu GluArg Thr Arg Leu Met Gly Tyr Trp Phe Val His Trp Phe Asp Gly Lys Leu Phe HisLeu Arg Ile Lys Ala Gly Gly Pro Asn Val Asp Gly Glu His Arg Ala Ile Arg ThrAla Glu His Pro Trp Leu Leu Arg Ala Arg Leu Asp Asp Ala Leu Glu Glu Ala LeuPro Lys Tyr Ala Ala Val Lys Lys Arg Pro Phe Thr Phe Leu Ala Gln Lys Asp GluLeu Ile Asp Ala Ala Ala Thr Ala Ala Gly Leu Ser His Arg Leu Leu Asn Ser PheLys Val Ile Pro Arg Phe Ala Leu Ser Pro Lys Ile Tyr Glu Pro Val Asp Gly ThrThr Arg Val Gly Val Phe Val Thr Ile Gly Met Arg Tyr Asp Ile Glu Ala Ser LeuArg Asp Leu Leu Glu Ala Gly Ile Asp Leu Arg Gly Met Tyr Val Val Arg Arg LysArg Gln Pro Gly Glu Arg Gly Leu Leu Gly Arg Val Arg Ala Ile Ser Asp Asp MetVal Gln Leu Phe Glu Glu Thr Asp Leu Ala Ser Val Asn Val Asn Asp Ala Lys LeuGlu Gly Ser Lys Glu Asn Phe Thr Arg Cys Leu Ser Ala Leu Leu Gly His Asn TyrLys Lys Leu Leu Asn Ala Leu Asp Asp Gln Glu Ala Gly Tyr Arg Thr Gly Pro ArgPhe Asp Asp Ala Val Arg Arg Met Gly Glu Phe Leu Ala Lys Lys Pro Ile Arg LeuAla Asp Asn Ile Asn Ala Gln Val Gly Asp Arg Ile Val Phe Ser Asn Glu Gly GlnAla Arg Asn Val Arg Leu Ala Pro Lys Val Glu Tyr Val Phe Asp Arg Thr Gly AlaLys Ser Ala Glu Tyr Ala Trp Arg Gly Leu Ser Gln Phe Gly Pro Phe Asp Arg ProSer Phe Ala Asn Arg Ser Pro Arg Ile Leu Val Val Tyr Pro Ser Ser Thr Gln GlyLys Val Glu Asn Phe Leu Ser Ala Phe Arg Asp Gly Met Gly Ser Asn Tyr Ser GlyPhe Ser Lys Gly Phe Val Asp Leu Met Gly Leu Thr Lys Val Glu Phe Val Met CysPro Val Glu Val Ser Ser Ala Asp Arg Asn Gly Ala His Thr Lys Tyr Asn Ser AlaIle Glu Asp Lys Leu Ala Gly Ala Gly Glu Val His Ala Gly Ile Val Val Leu PheGlu Asp His Ala Arg Leu Pro Asp Asp Arg Asn Pro Tyr Ile His Thr Lys Ser LeuLeu Leu Thr Leu Gly Val Pro Thr Gln Gln Val Arg Met Pro Thr Val Leu Leu GluPro Lys Ser Leu Gln Tyr Thr Leu Gln Asn Phe Ser Ile Ala Thr Tyr Ala Lys LeuAsn Gly Thr Pro Trp Thr Val Asn His Asp Lys Ala Ile Asn Asp Glu Leu Val ValGly Met Gly Leu Ala Glu Leu Ser Gly Ser Arg Thr Glu Lys Arg Gln Arg Phe ValGly Ile Thr Thr Val Phe Ala Gly Asp Gly Ser Tyr Leu Leu Gly Asn Val Ser LysGlu Cys Glu Tyr Glu Gly Tyr Ser Asp Ala Ile Arg Glu Ser Met Thr Gly Ile LeuArg Glu Leu Lys Lys Arg Asn Asn Trp Arg Pro Gly Asp Thr Val Arg Val Val PheHis Ala His Arg Pro Leu Lys Arg Val Asp Val Ala Ser Ile Val Phe Glu Cys ThrArg Glu Ile Gly Ser Asp Gln Asn Ile Gln Met Ala Phe Val Thr Val Ser His AspHis Pro Phe Val Leu Ile Asp Arg Ser Glu Arg Gly Leu Glu Ala Tyr Lys Gly SerThr Ala Arg Lys Gly Val Phe Ala Pro Pro Arg Gly Ala Ile Ser Arg Val Gly ArgLeu Thr Arg Leu Leu Ala Val Asn Ser Pro Gln Leu Ile Lys Arg Ala Asn Thr ProLeu Pro Thr Pro Leu Leu Val Ser Leu His Pro Asp Ser Thr Phe Lys Asp Val AspTyr Leu Ala Glu Gln Ala Leu Lys Phe Thr Ser Leu Ser Trp Arg Ser Thr Leu ProAla Ala Thr Pro Val Thr Ile Phe Tyr Ser Glu Arg Ile Ala Glu Leu Leu Gly ArgLeu Lys Ser Ile Pro Asn Trp Ser Ser Ala Asn Leu Asn Ile Lys Leu Lys Trp SerArg Trp Phe Leu Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Ser Gln Leu ValLys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys Tyr Val ProHis Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile LeuGlu Met Lys Val Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His LeuGly Gly Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp TyrGly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly GlnAla Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His Ile AsnPro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr Glu Phe Lys Phe Leu PheVal Ser Gly His Phe Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His IleThr Asn Cys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu MetIle Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly GluIle Asn Phe；

或者，构建得到的RsAgoE型的嵌合蛋白pAgoE的氨基酸序列为：Met Ala Pro Lys LysLys Arg Lys Val Gly Ile His Gly Val Pro Ala Ala Met Ala Pro Val Gln Ala AlaAsp Glu Met Tyr Asp Ser Asn Pro His Pro Asp Arg Arg Gln Leu Val Ser Asn GlyPhe Glu Val Asn Leu Pro Asp Gln Val Glu Val Ile Val Arg Asp Leu Pro Asp ProSer Lys Val Lys Glu Glu Arg Thr Arg Leu Met Gly Tyr Trp Phe Val His Trp PheAsp Gly Lys Leu Phe His Leu Arg Ile Lys Ala Gly Gly Pro Asn Val Asp Gly GluHis Arg Ala Ile Arg Thr Ala Glu His Pro Trp Leu Leu Arg Ala Arg Leu Asp AspAla Leu Glu Glu Ala Leu Pro Lys Tyr Ala Ala Val Lys Lys Arg Pro Phe Thr PheLeu Ala Gln Lys Asp Glu Leu Ile Asp Ala Ala Ala Thr Ala Ala Gly Leu Ser HisArg Leu Leu Asn Ser Phe Lys Val Ile Pro Arg Phe Ala Leu Ser Pro Lys Ile TyrGlu Pro Val Asp Gly Thr Thr Arg Val Gly Val Phe Val Thr Ile Gly Met Arg TyrAsp Ile Glu Ala Ser Leu Arg Asp Leu Leu Glu Ala Gly Ile Asp Leu Arg Gly MetTyr Val Val Arg Arg Lys Arg Gln Pro Gly Glu Arg Gly Leu Leu Gly Arg Val ArgAla Ile Ser Asp Asp Met Val Gln Leu Phe Glu Glu Thr Asp Leu Ala Ser Val AsnVal Asn Asp Ala Lys Leu Glu Gly Ser Lys Glu Asn Phe Thr Arg Cys Leu Ser AlaLeu Leu Gly His Asn Tyr Lys Lys Leu Leu Asn Ala Leu Asp Asp Gln Glu Ala GlyTyr Arg Thr Gly Pro Arg Phe Asp Asp Ala Val Arg Arg Met Gly Glu Phe Leu AlaLys Lys Pro Ile Arg Leu Ala Asp Asn Ile Asn Ala Gln Val Gly Asp Arg Ile ValPhe Ser Asn Glu Gly Gln Ala Arg Asn Val Arg Leu Ala Pro Lys Val Glu Tyr ValPhe Asp Arg Thr Gly Ala Lys Ser Ala Glu Tyr Ala Trp Arg Gly Leu Ser Gln PheGly Pro Phe Asp Arg Pro Ser Phe Ala Asn Arg Ser Pro Arg Ile Leu Val Val TyrPro Ser Ser Thr Gln Gly Lys Val Glu Asn Phe Leu Ser Ala Phe Arg Asp Gly MetGly Ser Asn Tyr Ser Gly Phe Ser Lys Gly Phe Val Asp Leu Met Gly Leu Thr LysVal Glu Phe Val Met Cys Pro Val Glu Val Ser Ser Ala Asp Arg Asn Gly Ala HisThr Lys Tyr Asn Ser Ala Ile Glu Asp Lys Leu Ala Gly Ala Gly Glu Val His AlaGly Ile Val Val Leu Phe Glu Asp His Ala Arg Leu Pro Asp Asp Arg Asn Pro TyrIle His Thr Lys Ser Leu Leu Leu Thr Leu Gly Val Pro Thr Gln Gln Val Arg MetPro Thr Val Leu Leu Glu Pro Lys Ser Leu Gln Tyr Thr Leu Gln Asn Phe Ser IleAla Thr Tyr Ala Lys Leu Asn Gly Thr Pro Trp Thr Val Asn His Asp Lys Ala IleAsn Asp Glu Leu Val Val Gly Met Gly Leu Ala Glu Leu Ser Gly Ser Arg Thr GluLys Arg Gln Arg Phe Val Gly Ile Thr Thr Val Phe Ala Gly Asp Gly Ser Tyr LeuLeu Gly Asn Val Ser Lys Glu Cys Glu Tyr Glu Gly Tyr Ser Asp Ala Ile Arg GluSer Met Thr Gly Ile Leu Arg Glu Leu Lys Lys Arg Asn Asn Trp Arg Pro Gly AspThr Val Arg Val Val Phe His Ala His Arg Pro Leu Lys Arg Val Asp Val Ala SerIle Val Phe Glu Cys Thr Arg Glu Ile Gly Ser Asp Gln Asn Ile Gln Met Ala PheVal Thr Val Ser His Asp His Pro Phe Val Leu Ile Asp Arg Ser Glu Arg Gly LeuGlu Ala Tyr Lys Gly Ser Thr Ala Arg Lys Gly Val Phe Ala Pro Pro Arg Gly AlaIle Ser Arg Val Gly Arg Leu Thr Arg Leu Leu Ala Val Asn Ser Pro Gln Leu IleLys Arg Ala Asn Thr Pro Leu Pro Thr Pro Leu Leu Val Ser Leu His Pro Asp SerThr Phe Lys Asp Val Asp Tyr Leu Ala Glu Gln Ala Leu Lys Phe Thr Ser Leu SerTrp Arg Ser Thr Leu Pro Ala Ala Thr Pro Val Thr Ile Phe Tyr Ser Glu Arg IleAla Glu Leu Leu Gly Arg Leu Lys Ser Ile Pro Asn Trp Ser Ser Ala Asn Leu AsnIle Lys Leu Lys Trp Ser Arg Trp Phe Leu Gly Gly Gly Gly Ser Gly Gly Gly GlySer Ser Ser Gln Leu Val Lys Ser Glu Leu Glu Glu Lys Lys Ser Glu Leu Arg HisLys Leu Lys Tyr Val Pro His Glu Tyr Ile Glu Leu Ile Glu Ile Ala Arg Asn SerThr Gln Asp Arg Ile Leu Glu Met Lys Val Met Glu Phe Phe Met Lys Val Tyr GlyTyr Arg Gly Lys His Leu Gly Gly Ser Arg Lys Pro Asp Gly Ala Ile Tyr Thr ValGly Ser Pro Ile Asp Tyr Gly Val Ile Val Asp Thr Lys Ala Tyr Ser Gly Gly TyrAsn Leu Pro Ile Gly Gln Ala Asp Glu Met Gln Arg Tyr Val Glu Glu Asn Gln ThrArg Asn Lys His Ile Asn Pro Asn Glu Trp Trp Lys Val Tyr Pro Ser Ser Val ThrGlu Phe Lys Phe Leu Phe Val Ser Gly His Phe Lys Gly Asn Tyr Lys Ala Gln LeuThr Arg Leu Asn His Ile Thr Asn Cys Asn Gly Ala Val Leu Ser Val Glu Glu LeuLeu Ile Gly Gly Glu Met Ile Lys Ala Gly Thr Leu Thr Leu Glu Glu Val Arg ArgLys Phe Asn Asn Gly Glu Ile Asn Phe；

其中，构建得到的MpAgoE型的嵌合蛋白pAgoE的氨基酸序列为：Met Ala Pro Lys LysLys Arg Lys Val Gly Ile His Gly Val Pro Ala Ala Ser Ser Gln Leu Val Lys SerGlu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys Tyr Val Pro His GluTyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu MetLys Val Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly GlySer Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly ValIle Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln Ala AspGlu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His Ile Asn Pro AsnGlu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val SerGly His Phe Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr AsnCys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile LysAla Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile AsnPhe Gly Gly Gly Gly Ser Met Tyr Leu Asn Leu Tyr Lys Ile Asp Ile Pro Lys LysIle Lys Arg Leu Tyr Phe Tyr Asn Pro Asp Met Glu Pro Lys Leu Phe Ala Arg AsnLeu Ser Arg Val Asn Asn Phe Lys Phe Gln Asp Ser Asn Asp Leu Val Trp Ile GluIle Pro Asp Ile Asp Phe Gln Ile Thr Pro Lys Asn Val Phe Gln Tyr Lys Val GluLys Glu Glu Ile Ile Lys Glu Glu Glu Asp Lys Lys Leu Phe Val Lys Thr Leu TyrLys Tyr Ile Lys Lys Leu Phe Leu Asp Asn Asp Phe Tyr Phe Lys Lys Gly Asn AsnPhe Ile Ser Asn Ser Glu Val Phe Ser Leu Asp Ser Asn Glu Asn Val Asn Ala HisLeu Thr Tyr Lys Ile Lys Ile His Asn Ile Ser Asn Glu Tyr Tyr Leu Ser Ile LeuPro Lys Phe Thr Phe Leu Ser Lys Glu Pro Ala Leu Glu Ser Ala Ile Lys Ser GlyTyr Leu Tyr Asn Ile Lys Ser Gly Lys Ser Phe Pro Tyr Ile Ser Gly Leu Asp GlyIle Leu Lys Ile Asp Ile Gly Asn Asn Gln Ile Val Glu Val Ala Tyr Pro Glu AsnTyr Leu Phe Asn Phe Thr Thr Arg Asp Ala Glu Lys Tyr Gly Phe Ser Lys Glu ValHis Glu Ile Tyr Lys Asn Lys Val Phe Glu Gly Phe Lys Lys Ile Pro Lys Thr LeuGly Phe Leu Asn Lys Ile Thr Asn Leu Asn Glu Asn Tyr Gln Leu Lys Asp Gly TyrLys Ile Phe Ile Asn Val Ile Tyr Lys Phe Lys Asn Gly Glu Ser Arg Tyr Ala LysAsp Val Phe Lys Tyr Ser Phe Tyr Lys Asn Glu Gln Pro Leu Lys Ala Ile Phe PhePhe Ser Ser Lys Lys Gln Phe Phe Glu Val Gln Lys Ser Leu Lys Glu Leu Phe HisAsn Lys His Ser Val Phe Tyr Arg Ala Ala Ala Glu Leu Gly Phe Ser Lys Val GluPhe Leu Arg Asp Ser Lys Thr Lys Ser Ser Ala Phe Leu Tyr Asn Pro Glu Glu PheThr Val Lys Asn Thr Glu Phe Ile Asn Gln Ile Glu Asp Asn Val Met Ala Ile ValLeu Leu Asp Lys Tyr Ile Gly Asn Ile Asp Pro Leu Val Arg Asn Phe Pro Asp AsnLeu Ile Leu Gln Pro Ile Leu Lys Glu Lys Leu Glu Asp Ile Lys Pro Phe Ile IleLys Ser Tyr Val Tyr Lys Met Gly Asn Phe Ile Pro Glu Cys Lys Pro Phe Ile LeuLys Lys Met Glu Asp Lys Glu Lys Asn Leu Tyr Ile Gly Ile Asp Leu Ser His AspThr Tyr Ala Arg Lys Thr Asn Leu Cys Ile Ala Ala Val Asp Asn Thr Gly Asp IleLeu Tyr Ile Gly Lys His Lys Asn Leu Glu Leu Asn Glu Lys Met Asn Leu Asp IleLeu Glu Lys Glu Tyr Ile Lys Ala Phe Glu Lys Tyr Ile Glu Lys Phe Asn Val SerPro Glu Asn Val Phe Ile Leu Arg Asp Gly Arg Phe Ile Glu Asp Ile Glu Ile IleLys Asn Phe Ile Ser Tyr Asn Asp Thr Lys Tyr Thr Leu Val Glu Val Asn Lys AsnThr Asn Ile Asn Ser Tyr Asp Asp Leu Lys Glu Trp Ile Ile Lys Leu Asp Glu AsnThr Tyr Ile Tyr Tyr Pro Lys Thr Phe Leu Asn Gln Lys Gly Val Glu Val Lys IleLeu Glu Asn Asn Thr Asp Tyr Thr Ile Glu Glu Ile Ile Glu Gln Ile Tyr Leu LeuThr Arg Val Ala His Ser Thr Pro Tyr Thr Asn Tyr Lys Leu Pro Tyr Pro Leu HisIle Ala Asn Lys Val Ala Leu Thr Asp Tyr Glu Trp Lys Leu Tyr Ile Pro Tyr；

其中，构建得到的dMpAgoE型的嵌合蛋白pAgoE的氨基酸序列为：Met Ala Pro Lys LysLys Arg Lys Val Gly Ile His Gly Val Pro Ala Ala Ser Ser Gln Leu Val Lys SerGlu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys Tyr Val Pro His GluTyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu MetLys Val Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly GlySer Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly ValIle Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln Ala AspGlu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His Ile Asn Pro AsnGlu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val SerGly His Phe Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr AsnCys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile LysAla Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile AsnPhe Gly Gly Gly Gly Ser Met Tyr Leu Asn Leu Tyr Lys Ile Asp Ile Pro Lys LysIle Lys Arg Leu Tyr Phe Tyr Asn Pro Asp Met Glu Pro Lys Leu Phe Ala Arg AsnLeu Ser Arg Val Asn Asn Phe Lys Phe Gln Asp Ser Asn Asp Leu Val Trp Ile GluIle Pro Asp Ile Asp Phe Gln Ile Thr Pro Lys Asn Val Phe Gln Tyr Lys Val GluLys Glu Glu Ile Ile Lys Glu Glu Glu Asp Lys Lys Leu Phe Val Lys Thr Leu TyrLys Tyr Ile Lys Lys Leu Phe Leu Asp Asn Asp Phe Tyr Phe Lys Lys Gly Asn AsnPhe Ile Ser Asn Ser Glu Val Phe Ser Leu Asp Ser Asn Glu Asn Val Asn Ala HisLeu Thr Tyr Lys Ile Lys Ile His Asn Ile Ser Asn Glu Tyr Tyr Leu Ser Ile LeuPro Lys Phe Thr Phe Leu Ser Lys Glu Pro Ala Leu Glu Ser Ala Ile Lys Ser GlyTyr Leu Tyr Asn Ile Lys Ser Gly Lys Ser Phe Pro Tyr Ile Ser Gly Leu Asp GlyIle Leu Lys Ile Asp Ile Gly Asn Asn Gln Ile Val Glu Val Ala Tyr Pro Glu AsnTyr Leu Phe Asn Phe Thr Thr Arg Asp Ala Glu Lys Tyr Gly Phe Ser Lys Glu ValHis Glu Ile Tyr Lys Asn Lys Val Phe Glu Gly Phe Lys Lys Ile Pro Lys Thr LeuGly Phe Leu Asn Lys Ile Thr Asn Leu Asn Glu Asn Tyr Gln Leu Lys Asp Gly TyrLys Ile Phe Ile Asn Val Ile Tyr Lys Phe Lys Asn Gly Glu Ser Arg Tyr Ala LysAsp Val Phe Lys Tyr Ser Phe Tyr Lys Asn Glu Gln Pro Leu Lys Ala Ile Phe PhePhe Ser Ser Lys Lys Gln Phe Phe Glu Val Gln Lys Ser Leu Lys Glu Leu Phe HisAsn Lys His Ser Val Phe Tyr Arg Ala Ala Ala Glu Leu Gly Phe Ser Lys Val GluPhe Leu Arg Asp Ser Lys Thr Lys Ser Ser Ala Phe Leu Tyr Asn Pro Glu Glu PheThr Val Lys Asn Thr Glu Phe Ile Asn Gln Ile Glu Asp Asn Val Met Ala Ile ValLeu Leu Asp Lys Tyr Ile Gly Asn Ile Asp Pro Leu Val Arg Asn Phe Pro Asp AsnLeu Ile Leu Gln Pro Ile Leu Lys Glu Lys Leu Glu Asp Ile Lys Pro Phe Ile IleLys Ser Tyr Val Tyr Lys Met Gly Asn Phe Ile Pro Glu Cys Lys Pro Phe Ile LeuLys Lys Met Glu Asp Lys Glu Lys Asn Leu Tyr Ile Gly Ile Ala Leu Ser His AspThr Tyr Ala Arg Lys Thr Asn Leu Cys Ile Ala Ala Val Asp Asn Thr Gly Asp IleLeu Tyr Ile Gly Lys His Lys Asn Leu Glu Leu Asn Glu Lys Met Asn Leu Asp IleLeu Glu Lys Glu Tyr Ile Lys Ala Phe Glu Lys Tyr Ile Glu Lys Phe Asn Val SerPro Glu Asn Val Phe Ile Leu Arg Asp Gly Arg Phe Ile Glu Asp Ile Glu Ile IleLys Asn Phe Ile Ser Tyr Asn Asp Thr Lys Tyr Thr Leu Val Glu Val Asn Lys AsnThr Asn Ile Asn Ser Tyr Asp Asp Leu Lys Glu Trp Ile Ile Lys Leu Asp Glu AsnThr Tyr Ile Tyr Tyr Pro Lys Thr Phe Leu Asn Gln Lys Gly Val Glu Val Lys IleLeu Glu Asn Asn Thr Asp Tyr Thr Ile Glu Glu Ile Ile Glu Gln Ile Tyr Leu LeuThr Arg Val Ala His Ser Thr Pro Tyr Thr Asn Tyr Lys Leu Pro Tyr Pro Leu HisIle Ala Asn Lys Val Ala Leu Thr Asp Tyr Glu Trp Lys Leu Tyr Ile Pro Tyr；

或者，构建得到的dMpAgoE型的嵌合蛋白pAgoE的氨基酸序列为：Met Ala Pro Lys LysLys Arg Lys Val Gly Ile His Gly Val Pro Ala Ala Ser Ser Gln Leu Val Lys SerGlu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys Tyr Val Pro His GluTyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu MetLys Val Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly GlySer Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly ValIle Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln Ala AspGlu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His Ile Asn Pro AsnGlu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val SerGly His Phe Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr AsnCys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile LysAla Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile AsnPhe Gly Gly Gly Gly Ser Met Tyr Leu Asn Leu Tyr Lys Ile Asp Ile Pro Lys LysIle Lys Arg Leu Tyr Phe Tyr Asn Pro Asp Met Glu Pro Lys Leu Phe Ala Arg AsnLeu Ser Arg Val Asn Asn Phe Lys Phe Gln Asp Ser Asn Asp Leu Val Trp Ile GluIle Pro Asp Ile Asp Phe Gln Ile Thr Pro Lys Asn Val Phe Gln Tyr Lys Val GluLys Glu Glu Ile Ile Lys Glu Glu Glu Asp Lys Lys Leu Phe Val Lys Thr Leu TyrLys Tyr Ile Lys Lys Leu Phe Leu Asp Asn Asp Phe Tyr Phe Lys Lys Gly Asn AsnPhe Ile Ser Asn Ser Glu Val Phe Ser Leu Asp Ser Asn Glu Asn Val Asn Ala HisLeu Thr Tyr Lys Ile Lys Ile His Asn Ile Ser Asn Glu Tyr Tyr Leu Ser Ile LeuPro Lys Phe Thr Phe Leu Ser Lys Glu Pro Ala Leu Glu Ser Ala Ile Lys Ser GlyTyr Leu Tyr Asn Ile Lys Ser Gly Lys Ser Phe Pro Tyr Ile Ser Gly Leu Asp GlyIle Leu Lys Ile Asp Ile Gly Asn Asn Gln Ile Val Glu Val Ala Tyr Pro Glu AsnTyr Leu Phe Asn Phe Thr Thr Arg Asp Ala Glu Lys Tyr Gly Phe Ser Lys Glu ValHis Glu Ile Tyr Lys Asn Lys Val Phe Glu Gly Phe Lys Lys Ile Pro Lys Thr LeuGly Phe Leu Asn Lys Ile Thr Asn Leu Asn Glu Asn Tyr Gln Leu Lys Asp Gly TyrLys Ile Phe Ile Asn Val Ile Tyr Lys Phe Lys Asn Gly Glu Ser Arg Tyr Ala LysAsp Val Phe Lys Tyr Ser Phe Tyr Lys Asn Glu Gln Pro Leu Lys Ala Ile Phe PhePhe Ser Ser Lys Lys Gln Phe Phe Glu Val Gln Lys Ser Leu Lys Glu Leu Phe HisAsn Lys His Ser Val Phe Tyr Arg Ala Ala Ala Glu Leu Gly Phe Ser Lys Val GluPhe Leu Arg Asp Ser Lys Thr Lys Ser Ser Ala Phe Leu Tyr Asn Pro Glu Glu PheThr Val Lys Asn Thr Glu Phe Ile Asn Gln Ile Glu Asp Asn Val Met Ala Ile ValLeu Leu Asp Lys Tyr Ile Gly Asn Ile Asp Pro Leu Val Arg Asn Phe Pro Asp AsnLeu Ile Leu Gln Pro Ile Leu Lys Glu Lys Leu Glu Asp Ile Lys Pro Phe Ile IleLys Ser Tyr Val Tyr Lys Met Gly Asn Phe Ile Pro Glu Cys Lys Pro Phe Ile LeuLys Lys Met Glu Asp Lys Glu Lys Asn Leu Tyr Ile Gly Ile Ala Leu Ser His AspThr Tyr Ala Arg Lys Thr Asn Leu Cys Ile Ala Ala Val Asp Asn Thr Gly Asp IleLeu Tyr Ile Gly Lys His Lys Asn Leu Glu Leu Asn Glu Lys Met Asn Leu Asp IleLeu Glu Lys Glu Tyr Ile Lys Ala Phe Glu Lys Tyr Ile Glu Lys Phe Asn Val SerPro Glu Asn Val Phe Ile Leu Arg Asp Gly Arg Phe Ile Glu Asp Ile Glu Ile IleLys Asn Phe Ile Ser Tyr Asn Asp Thr Lys Tyr Thr Leu Val Glu Val Asn Lys AsnThr Asn Ile Asn Ser Tyr Asp Asp Leu Lys Glu Trp Ile Ile Lys Leu Asp Glu AsnThr Tyr Ile Tyr Tyr Pro Lys Thr Phe Leu Asn Gln Lys Gly Val Glu Val Lys IleLeu Glu Asn Asn Thr Asp Tyr Thr Ile Glu Glu Ile Ile Glu Gln Ile Tyr Leu LeuThr Arg Val Ala His Ser Thr Pro Tyr Thr Asn Tyr Lys Leu Pro Tyr Pro Leu HisIle Ala Asn Lys Val Ala Leu Thr Asp Tyr Glu Trp Lys Leu Tyr Ile Pro Tyr GlyGly Gly Gly Ser Gly Gly Gly Gly Ser Ser Ser Gln Leu Val Lys Ser Glu Leu GluGlu Lys Lys Ser Glu Leu Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr Ile GluLeu Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys Val MetGlu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser Arg LysPro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile Val AspThr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln Ala Asp Glu Met GlnArg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His Ile Asn Pro Asn Glu Trp TrpLys Val Tyr Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly His PheLys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys Asn GlyAla Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala Gly ThrLeu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile Asn Phe；

其中，构建得到的dNgAgoE型的嵌合蛋白pAgoE的氨基酸序列为：Met Ala Pro Lys LysLys Arg Lys Val Gly Ile His Gly Val Pro Ala Ala Ser Ser Gln Leu Val Lys SerGlu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys Tyr Val Pro His GluTyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu MetLys Val Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly GlySer Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly ValIle Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln Ala AspGlu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His Ile Asn Pro AsnGlu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val SerGly His Phe Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr AsnCys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile LysAla Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile AsnPhe Gly Gly Gly Gly Ser Met Thr Val Ile Asp Leu Asp Ser Thr Thr Thr Ala AspGlu Leu Thr Ser Gly His Thr Tyr Asp Ile Ser Val Thr Leu Thr Gly Val Tyr AspAsn Thr Asp Glu Gln His Pro Arg Met Ser Leu Ala Phe Glu Gln Asp Asn Gly GluArg Arg Tyr Ile Thr Leu Trp Lys Asn Thr Thr Pro Lys Asp Val Phe Thr Tyr AspTyr Ala Thr Gly Ser Thr Tyr Ile Phe Thr Asn Ile Asp Tyr Glu Val Lys Asp GlyTyr Glu Asn Leu Thr Ala Thr Tyr Gln Thr Thr Val Glu Asn Ala Thr Ala Gln GluVal Gly Thr Thr Asp Glu Asp Glu Thr Phe Ala Gly Gly Glu Pro Leu Asp His HisLeu Asp Asp Ala Leu Asn Glu Thr Pro Asp Asp Ala Glu Thr Glu Ser Asp Ser GlyHis Val Met Thr Ser Phe Ala Ser Arg Asp Gln Leu Pro Glu Trp Thr Leu His ThrTyr Thr Leu Thr Ala Thr Asp Gly Ala Lys Thr Asp Thr Glu Tyr Ala Arg Arg ThrLeu Ala Tyr Thr Val Arg Gln Glu Leu Tyr Thr Asp His Asp Ala Ala Pro Val AlaThr Asp Gly Leu Met Leu Leu Thr Pro Glu Pro Leu Gly Glu Thr Pro Leu Asp LeuAsp Cys Gly Val Arg Val Glu Ala Asp Glu Thr Arg Thr Leu Asp Tyr Thr Thr AlaLys Asp Arg Leu Leu Ala Arg Glu Leu Val Glu Glu Gly Leu Lys Arg Ser Leu TrpAsp Asp Tyr Leu Val Arg Gly Ile Asp Glu Val Leu Ser Lys Glu Pro Val Leu ThrCys Asp Glu Phe Asp Leu His Glu Arg Tyr Asp Leu Ser Val Glu Val Gly His SerGly Arg Ala Tyr Leu His Ile Asn Phe Arg His Arg Phe Val Pro Lys Leu Thr LeuAla Asp Ile Asp Asp Asp Asn Ile Tyr Pro Gly Leu Arg Val Lys Thr Thr Tyr ArgPro Arg Arg Gly His Ile Val Trp Gly Leu Arg Asp Glu Cys Ala Thr Asp Ser LeuAsn Thr Leu Gly Asn Gln Ser Val Val Ala Tyr His Arg Asn Asn Gln Thr Pro IleAsn Thr Asp Leu Leu Asp Ala Ile Glu Ala Ala Asp Arg Arg Val Val Glu Thr ArgArg Gln Gly His Gly Asp Asp Ala Val Ser Phe Pro Gln Glu Leu Leu Ala Val GluPro Asn Thr His Gln Ile Lys Gln Phe Ala Ser Asp Gly Phe His Gln Gln Ala ArgSer Lys Thr Arg Leu Ser Ala Ser Arg Cys Ser Glu Lys Ala Gln Ala Phe Ala GluArg Leu Asp Pro Val Arg Leu Asn Gly Ser Thr Val Glu Phe Ser Ser Glu Phe PheThr Gly Asn Asn Glu Gln Gln Leu Arg Leu Leu Tyr Glu Asn Gly Glu Ser Val LeuThr Phe Arg Asp Gly Ala Arg Gly Ala His Pro Asp Glu Thr Phe Ser Lys Gly IleVal Asn Pro Pro Glu Ser Phe Glu Val Ala Val Val Leu Pro Glu Gln Gln Ala AspThr Cys Lys Ala Gln Trp Asp Thr Met Ala Asp Leu Leu Asn Gln Ala Gly Ala ProPro Thr Arg Ser Glu Thr Val Gln Tyr Asp Ala Phe Ser Ser Pro Glu Ser Ile SerLeu Asn Val Ala Gly Ala Ile Asp Pro Ser Glu Val Asp Ala Ala Phe Val Val LeuPro Pro Asp Gln Glu Gly Phe Ala Asp Leu Ala Ser Pro Thr Glu Thr Tyr Asp GluLeu Lys Lys Ala Leu Ala Asn Met Gly Ile Tyr Ser Gln Met Ala Tyr Phe Asp ArgPhe Arg Asp Ala Lys Ile Phe Tyr Thr Arg Asn Val Ala Leu Gly Leu Leu Ala AlaAla Gly Gly Val Ala Phe Thr Thr Glu His Ala Met Pro Gly Asp Ala Asp Met PheIle Gly Ile Ala Val Ser Arg Ser Tyr Pro Glu Asp Gly Ala Ser Gly Gln Ile AsnIle Ala Ala Thr Ala Thr Ala Val Tyr Lys Asp Gly Thr Ile Leu Gly His Ser SerThr Arg Pro Gln Leu Gly Glu Lys Leu Gln Ser Thr Asp Val Arg Asp Ile Met LysAsn Ala Ile Leu Gly Tyr Gln Gln Val Thr Gly Glu Ser Pro Thr His Ile Val IleHis Arg Asp Gly Phe Met Asn Glu Asp Leu Asp Pro Ala Thr Glu Phe Leu Asn GluGln Gly Val Glu Tyr Asp Ile Val Glu Ile Arg Lys Gln Pro Gln Thr Arg Leu LeuAla Val Ser Asp Val Gln Tyr Asp Thr Pro Val Lys Ser Ile Ala Ala Ile Asn GlnAsn Glu Pro Arg Ala Thr Val Ala Thr Phe Gly Ala Pro Glu Tyr Leu Ala Thr ArgAsp Gly Gly Gly Leu Pro Arg Pro Ile Gln Ile Glu Arg Val Ala Gly Glu Thr AspIle Glu Thr Leu Thr Arg Gln Val Tyr Leu Leu Ser Gln Ser His Ile Gln Val HisAsn Ser Thr Ala Arg Leu Pro Ile Thr Thr Ala Tyr Ala Asp Gln Ala Ser Thr HisAla Thr Lys Gly Tyr Leu Val Gln Thr Gly Ala Phe Glu Ser Asn Val Gly Phe Leu；

或者，构建得到的dNgAgoE型的嵌合蛋白pAgoE的氨基酸序列为：Met Ala Pro Lys LysLys Arg Lys Val Gly Ile His Gly Val Pro Ala Ala Ser Ser Gln Leu Val Lys SerGlu Leu Glu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys Tyr Val Pro His GluTyr Ile Glu Leu Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu MetLys Val Met Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly GlySer Arg Lys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly ValIle Val Asp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln Ala AspGlu Met Gln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His Ile Asn Pro AsnGlu Trp Trp Lys Val Tyr Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val SerGly His Phe Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr AsnCys Asn Gly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile LysAla Gly Thr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile AsnPhe Gly Gly Gly Gly Ser Met Thr Val Ile Asp Leu Asp Ser Thr Thr Thr Ala AspGlu Leu Thr Ser Gly His Thr Tyr Asp Ile Ser Val Thr Leu Thr Gly Val Tyr AspAsn Thr Asp Glu Gln His Pro Arg Met Ser Leu Ala Phe Glu Gln Asp Asn Gly GluArg Arg Tyr Ile Thr Leu Trp Lys Asn Thr Thr Pro Lys Asp Val Phe Thr Tyr AspTyr Ala Thr Gly Ser Thr Tyr Ile Phe Thr Asn Ile Asp Tyr Glu Val Lys Asp GlyTyr Glu Asn Leu Thr Ala Thr Tyr Gln Thr Thr Val Glu Asn Ala Thr Ala Gln GluVal Gly Thr Thr Asp Glu Asp Glu Thr Phe Ala Gly Gly Glu Pro Leu Asp His HisLeu Asp Asp Ala Leu Asn Glu Thr Pro Asp Asp Ala Glu Thr Glu Ser Asp Ser GlyHis Val Met Thr Ser Phe Ala Ser Arg Asp Gln Leu Pro Glu Trp Thr Leu His ThrTyr Thr Leu Thr Ala Thr Asp Gly Ala Lys Thr Asp Thr Glu Tyr Ala Arg Arg ThrLeu Ala Tyr Thr Val Arg Gln Glu Leu Tyr Thr Asp His Asp Ala Ala Pro Val AlaThr Asp Gly Leu Met Leu Leu Thr Pro Glu Pro Leu Gly Glu Thr Pro Leu Asp LeuAsp Cys Gly Val Arg Val Glu Ala Asp Glu Thr Arg Thr Leu Asp Tyr Thr Thr AlaLys Asp Arg Leu Leu Ala Arg Glu Leu Val Glu Glu Gly Leu Lys Arg Ser Leu TrpAsp Asp Tyr Leu Val Arg Gly Ile Asp Glu Val Leu Ser Lys Glu Pro Val Leu ThrCys Asp Glu Phe Asp Leu His Glu Arg Tyr Asp Leu Ser Val Glu Val Gly His SerGly Arg Ala Tyr Leu His Ile Asn Phe Arg His Arg Phe Val Pro Lys Leu Thr LeuAla Asp Ile Asp Asp Asp Asn Ile Tyr Pro Gly Leu Arg Val Lys Thr Thr Tyr ArgPro Arg Arg Gly His Ile Val Trp Gly Leu Arg Asp Glu Cys Ala Thr Asp Ser LeuAsn Thr Leu Gly Asn Gln Ser Val Val Ala Tyr His Arg Asn Asn Gln Thr Pro IleAsn Thr Asp Leu Leu Asp Ala Ile Glu Ala Ala Asp Arg Arg Val Val Glu Thr ArgArg Gln Gly His Gly Asp Asp Ala Val Ser Phe Pro Gln Glu Leu Leu Ala Val GluPro Asn Thr His Gln Ile Lys Gln Phe Ala Ser Asp Gly Phe His Gln Gln Ala ArgSer Lys Thr Arg Leu Ser Ala Ser Arg Cys Ser Glu Lys Ala Gln Ala Phe Ala GluArg Leu Asp Pro Val Arg Leu Asn Gly Ser Thr Val Glu Phe Ser Ser Glu Phe PheThr Gly Asn Asn Glu Gln Gln Leu Arg Leu Leu Tyr Glu Asn Gly Glu Ser Val LeuThr Phe Arg Asp Gly Ala Arg Gly Ala His Pro Asp Glu Thr Phe Ser Lys Gly IleVal Asn Pro Pro Glu Ser Phe Glu Val Ala Val Val Leu Pro Glu Gln Gln Ala AspThr Cys Lys Ala Gln Trp Asp Thr Met Ala Asp Leu Leu Asn Gln Ala Gly Ala ProPro Thr Arg Ser Glu Thr Val Gln Tyr Asp Ala Phe Ser Ser Pro Glu Ser Ile SerLeu Asn Val Ala Gly Ala Ile Asp Pro Ser Glu Val Asp Ala Ala Phe Val Val LeuPro Pro Asp Gln Glu Gly Phe Ala Asp Leu Ala Ser Pro Thr Glu Thr Tyr Asp GluLeu Lys Lys Ala Leu Ala Asn Met Gly Ile Tyr Ser Gln Met Ala Tyr Phe Asp ArgPhe Arg Asp Ala Lys Ile Phe Tyr Thr Arg Asn Val Ala Leu Gly Leu Leu Ala AlaAla Gly Gly Val Ala Phe Thr Thr Glu His Ala Met Pro Gly Asp Ala Asp Met PheIle Gly Ile Ala Val Ser Arg Ser Tyr Pro Glu Asp Gly Ala Ser Gly Gln Ile AsnIle Ala Ala Thr Ala Thr Ala Val Tyr Lys Asp Gly Thr Ile Leu Gly His Ser SerThr Arg Pro Gln Leu Gly Glu Lys Leu Gln Ser Thr Asp Val Arg Asp Ile Met LysAsn Ala Ile Leu Gly Tyr Gln Gln Val Thr Gly Glu Ser Pro Thr His Ile Val IleHis Arg Asp Gly Phe Met Asn Glu Asp Leu Asp Pro Ala Thr Glu Phe Leu Asn GluGln Gly Val Glu Tyr Asp Ile Val Glu Ile Arg Lys Gln Pro Gln Thr Arg Leu LeuAla Val Ser Asp Val Gln Tyr Asp Thr Pro Val Lys Ser Ile Ala Ala Ile Asn GlnAsn Glu Pro Arg Ala Thr Val Ala Thr Phe Gly Ala Pro Glu Tyr Leu Ala Thr ArgAsp Gly Gly Gly Leu Pro Arg Pro Ile Gln Ile Glu Arg Val Ala Gly Glu Thr AspIle Glu Thr Leu Thr Arg Gln Val Tyr Leu Leu Ser Gln Ser His Ile Gln Val HisAsn Ser Thr Ala Arg Leu Pro Ile Thr Thr Ala Tyr Ala Asp Gln Ala Ser Thr HisAla Thr Lys Gly Tyr Leu Val Gln Thr Gly Ala Phe Glu Ser Asn Val Gly Phe LeuGly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ser Ser Gln Leu Val Lys Ser Glu LeuGlu Glu Lys Lys Ser Glu Leu Arg His Lys Leu Lys Tyr Val Pro His Glu Tyr IleGlu Leu Ile Glu Ile Ala Arg Asn Ser Thr Gln Asp Arg Ile Leu Glu Met Lys ValMet Glu Phe Phe Met Lys Val Tyr Gly Tyr Arg Gly Lys His Leu Gly Gly Ser ArgLys Pro Asp Gly Ala Ile Tyr Thr Val Gly Ser Pro Ile Asp Tyr Gly Val Ile ValAsp Thr Lys Ala Tyr Ser Gly Gly Tyr Asn Leu Pro Ile Gly Gln Ala Asp Glu MetGln Arg Tyr Val Glu Glu Asn Gln Thr Arg Asn Lys His Ile Asn Pro Asn Glu TrpTrp Lys Val Tyr Pro Ser Ser Val Thr Glu Phe Lys Phe Leu Phe Val Ser Gly HisPhe Lys Gly Asn Tyr Lys Ala Gln Leu Thr Arg Leu Asn His Ile Thr Asn Cys AsnGly Ala Val Leu Ser Val Glu Glu Leu Leu Ile Gly Gly Glu Met Ile Lys Ala GlyThr Leu Thr Leu Glu Glu Val Arg Arg Lys Phe Asn Asn Gly Glu Ile Asn Phe。