[发明专利]玉米螟谷胱甘肽-S-转移酶基因的cDNA及其RNAi干扰工程菌的构建

摘要

玉米螟谷胱甘肽-S-转移酶基因的cDNA及其RNAi干扰工程菌的构建，它涉及谷胱甘肽-S-转移酶基因的cDNA及其RNAi干扰工程菌构建。玉米螟谷胱甘肽-S-转移酶基因的核苷酸序列见SEQ ID No.1，氨基酸序列见SEQ ID No.2。构建：一、提取玉米螟总RNA，合成cDNA第一链，设计引物，进行PCR扩增，得Of-GST-RNAi320；二、Of-GST-RNAi320与L4440进行双酶切，回收所需目的片段，连接并构建L4440-Of-GST；三、将L4440-Of-GST转化到大肠杆菌HT115(DE3)中即完成。本发明操作简单、成本低，对玉米螟生长发育起到明显的抑制作用。

主权项

1.玉米螟谷胱甘肽-S-转移酶基因的cDNA，其特征在于玉米螟谷胱甘肽-S-转移酶基因的cDNA全长为886bp，核苷酸序列为：TCGCGGATCCACAGCCTACTGATGATCAGTCGATGGAAAAGTCTTACTGTGGAGTTCTGAAGCGGTGCACGCGAGTTTCAAAGCTACATCGATCGACCTTTACTACACGGCCGGGTCAGCGCCCTGTCGCCTGGTGTTACTGGTGGCTGCCGCTCTGAACCTCGAACTCAACCTTAAGCCTCTAGACTTAAGGGCTGGAGAACAGCTCACACCAGAATTTTTACAGCTGAACCCACAACACACAGTTCCCACGATCATAGACGACGGCTTTTCGCTAAATCCTCCACACACAGTTCCCACGATCGTAGACGACGGCTTTTCGCTATGGGAATCGCGCGCCATCAGCCGATACCTGGTCAACAAGTACGGAGGGGCAGCCCTGTACCCTGAGGACCCGAAGGCGAGGGCTTTGGTGGACCAGAGACTGGACTTCGACTTGGGTACACTGTACCCCAGATTTGCGACGTACTTTTATCCCCAAGTCCTCGCTGGTGCTCCTGCTGACGAAGCCTTGTTGAAGATGCTGGAAGAGGCTCTCAAGTTCCTGGACACCTTCCTCGAAGGTCAGAAGTATGCTGCGGGCTCAGAACTGACCTTGGCTGACCTGTCGCTCGTGGCTACTGTGTCTACTATTGACGCTGCTGGCATATCCATCGAATCCTACCAGAGTGTCAACAAGTGGTTCACGCTGGTCAAATCTACCGCCCCCAAATACGACGAAGCGAACGGCCAAGGCGTAGAAATGTTTAGAGCCTTCGTGGCACAGCTCAAAGCCAAGACGGAGCTGTCGTGAGACTGGTATAGTGCATAGTTTTGTATTAAGTAAATGTTTGATACATAAGTGAATTTGAAATTTAAAAGTAAAAAAAAAAAA；该基因所编码的氨基酸序列为：MetSerIleAspLeuTyrTyrThrAlaGlySerAlaProCysArgLeuValLeuLeuValAlaAlaAlaLeuAsnLeuGluLeuAsnLeuLysProLeuAspLeuArgAlaGlyGluGlnLeuThrProGluPheLeuGlnLeuAsnProGlnHisThrValProThrIleIleAspAspGlyPheSerLeuAsnProProHisThrValProThrIleValAspAspGlyPheSerLeuTrpGluSerArgAlaIleSerArgTyrLeuValAsnLysTyrGlyGlyAlaAlaLeuTyrProGluAspProLysAlaArgAlaLeuValAspGlnArgLeuAspPheAspLeuGlyThrLeuTyrProArgPheAlaThrTyrPheTyrProGlnValLeuAlaGlyAlaProAlaAspGluAlaLeuLeuLysMetLeuGluGluAlaLeuLysPheLeuAspThrPheLeuGluGlyGlnLysTyrAlaAlaGlySerGluLeuThrLeuAlaAspLeuSerLeuValAlaThrValSerThrIleAspAlaAlaGlyIleSerIleGluSerTyrGlnSerValAsnLysTrpPheThrLeuValLysSerThrAlaProLysTyrAspGluAlaAsnGly GlnGlyVal GluMetPheArgAlaPheValAlaGlnLeuLysAlaLysThrGluLeu。