抗菌肽Defensin基因pcr3.1真核表达载体的构建  

Construction of Eukaryotic Expression Vector of Antimicrobial Peptide Gene Defensin

作　　者：张守纯[1] 刘潇然[1] 郭宇航[1] 

机构地区：[1]沈阳农业大学畜牧兽医学院,沈阳110161

出　　处：《湖北农业科学》2009年第1期39-41,共3页Hubei Agricultural Sciences

摘　　要：通过克隆果蝇的defensin基因,构建了defensin基因的哺乳动物细胞真核表达载体。把黑腹果蝇基因组DNA通过PCR获得defensin,克隆载体pMD18-T/defensin,并将其转化到大肠杆菌里,最后进行抗菌肽基因重组真核表达载体pcr3.1/defensin的构建和鉴定。结果表明,以果蝇总DNA为模板扩增出280bp左右的特异性条带,测序结果与GeneBank测序结果相比,除一个碱基外,其余的完全一致,译成的氨基酸序列相同。对重组质粒pcr3.1/defensin进行双酶切鉴定并测序,结果也完全一致。所以,重组质粒pcr3.1/defensin由真核载体pcr3.1和defensin DNA片段组成。且插入的defensin DNA片段与已知序列完全相同。The eukaryotic expression vector of defensin was constructed according to the defensin gene of drosophila.De-fensin gene was obtained from drosophila genome DNA by PCR.Vector pMD18-T / defensin was cloned and transformed into Escherichia coli,and the recombinant pcr3.1 / defensin was constructed and identified.The result showed that 280 bp specif-ic fragment was amplified and identitied.Sequencing result showed that the reading frame was not changed except one base compared with the GeneBank,and amino acid sequence was the same.The result of identification and sequencing of pcr3.1 / defensin plasmid was the accordance with the expected length.

关 键 词：抗菌肽 defensin基因 PCR 真核表达载体构建 

分 类 号：Q516[生物学—生物化学] Q785