| 蜂毒溶血肽基因的定点诱变及其在大肠杆菌中的表达 |
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| 引用本文： | 王关林,李大力,方宏筠.蜂毒溶血肽基因的定点诱变及其在大肠杆菌中的表达[J].遗传学报,2000,27(2):176-182. |
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| 作者姓名： | 王关林 李大力 方宏筠 |
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| 作者单位： | [1]辽宁师范大学生物工程研究所,大连 [2]大连理工大学化工学院,大连 |
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| 摘 要： | 从蜜蜂毒腺中提取总ＲＮＡ，通过ＲＴ－ＰＣＲ方法扩增得到了蜂毒溶血肽前体蛋白的ｃＤＮＡ，再进一步通过定点诱变在蜂毒溶血肽序列前引入了羟胺裂解位点，构建了与β－半乳糖苷酶部分序列相融合的蜂毒溶血肽诱变蛋白表达载体，序列分析结果表明，成功地引入了目的密码子且与β－半乳糖苷酶部分序列构成正确的读码框，并在大肠杆菌中表达了诱变蛋白，为基因工程生产蜂毒溶血肽提供了新途径。
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| 关 键 词： | 蜂毒溶血肽 定点诱变 大肠杆菌 基因表达 |
Site-sirected Mutagenesis of Melittin Gene and Its Expression in Escherichia coli |
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| WANG Guan-Lin,LI Da-Li,FANG Hong-Jun.Site-sirected Mutagenesis of Melittin Gene and Its Expression in Escherichia coli[J].Journal of Genetics and Genomics,2000,27(2):176-182. |
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| Authors: | WANG Guan-Lin LI Da-Li FANG Hong-Jun |
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| Abstract: | The cDNA encoding promelittin was obtained from the total RNA of bee poison gland by RT-PCR Moreover, hydroxylamine clearage site was arranged before the melittin sequences by site-directed mutagenesis. The expression vector containing the mutagenic promelittin sequence with partial sequence of B -galactosidase was constructed. The result of DNA sequence analysis demonstrated that the obtained cDNA sequence include the desired codon and the reading frame of fusion gene was correct. The induced protein was expressed in Escherichia coli |
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| Keywords: | melittin mutagenesis expression |
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