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皮蝇素A基因的克隆与序列分析
Molecular Cloning and Sequence Analysis of Hypodermin A Gene of Hypoderma Species
【摘要】 首先提取采自甘肃玛曲的皮蝇一期幼虫的总RNA,合成cDNA,根据设计的特异引物,利用PCR技术扩增出Hypodermin A(HA)基因片段,将此片段与PGEM-T Easy载体连接,构建克隆载体PGEM-HA,转化大肠杆菌JM109,筛选阳性克隆,进行测序与分析。结果表明,扩增的HA基因长678bp,编码229氨基酸。克隆到的基因序列与NCBI GenBank上登陆的皮蝇HA基因序列核甘酸同源性为96.6%,推导氨基酸同源性达97.8%。该基因的成功克隆为其进一步表达打下了坚实的基础。
【Abstract】 In this thesis,the total RNA of first-stage larvae from Hypoderma species colleted from Maqu Gansu was isolated and used for synthesis of cDNA.According to specific primer,HA gene was amplified with PCR and ligated into PEGM-T Easy vector.The recombinants transformed into E.coli. Positive recombinants were selected,sequenced and analyzed.The cloned gene is 678 bp in length,encodes 229 aa.In comparison with NCBI GenBank data,the homologies of the nucleotide sequence is 96.6% and of amino sequence is 97.8%.This has laid a foundation for expression of HA gene.
【Key words】 Hypoderma species; Hypodermin A; cloning; sequencing;
- 【文献出处】 畜牧兽医杂志 ,Journal of Animal Science and Veterinary Medicine , 编辑部邮箱 ,2007年06期
- 【分类号】S852.7
- 【被引频次】1
- 【下载频次】52