›› 1998, Vol. 41 ›› Issue (3): 225-230.

• 研究论文 •    下一篇

用低G/C%含量引物通过PCR扩增家蝇细胞色素P—450 cDNA

康巧华 陈年春 周顺伍 齐顺章   

  • 出版日期:1998-08-20 发布日期:1998-08-20

AMPLIFICATION OF HOUSEFLY P-450 cDNA BY PCR DIRECTED BY A PAIR OF LOW G/C CONTENT PRIMERS

Kang Qiaohua Chen Nianchun Zhou Shunwu Qi Shunzhang   

  • Online:1998-08-20 Published:1998-08-20

摘要: 根据昆虫细胞色素P-450基因的多型性和遗传多态性,以苯巴比妥钠诱导、室内饲养的杀虫剂敏感种群雌性家蝇Musca omestica vicina Macquart为材料,提取总RNA,以0ligo(dT)-纤维素亲和层析分离出总mRNA;以此为模板反转录合成总cDNA。再以总cDNA为模板,以P-450CYP6A1cDNA序列为参考设计一对低G/C%含量引物,进行PCR扩增,获得1.5kb左右的预期目的片段。

关键词: 家蝇, 细胞色素P—450, cDNA, PCR, 遗传多态性

Abstract: Based on multiplicity and genetic polymorphism of P-450 genes, total MrNA was isolated through oligo (Dt) -cellulose column from total RNAs of an insecticidesusceptible strain of the housefly, Musca dornestica vicina Macquart treated with sodium phenobarbital in the drinking water. Double-stranded Cdna were synthesized by reverse-transcription and used as template for polymerase chain reaction(PCR). The 1.5kb long anticipated DNA fragment coding for P-450 protein was amplified by PCR,using a pair of low G/C% content primers designed according to CYP6A1 Cdna gene.

Key words: housefly, cytochrome P-450, cDNA, PCR, genetic polymorphism