用低G/C%含量引物通过PCR扩增家蝇细胞色素P-450 cDNA  

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

作　　者：康巧华[1] 陈年春[1] 周顺伍[2,3] 齐顺章[2,3] 

机构地区：[1]中国农业大学应用化学系 [2]中国农业大学生物学院 [3]北京市农林科学院植保环保研究所

出　　处：《昆虫学报》1998年第3期225-230,共6页Acta Entomologica Sinica

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ＭｕｓｃａｄｏｍｅｓｔｉｃａｖｉｃｉｎａＭａｃｑｕａｒｔ为材料，提取总ＲＮＡ，以Ｏｌｉｇｏ（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。Based on multiplicity and genetic polymorphism of P450 genes, total mRNA was isolated through oligo(dT)cellulose column from total RNAs of an insecticidesusceptible strain of the housefly, Musca domestica 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.

关 键 词：家蝇 细胞色素P-450 CDNA PCR 遗传多态性 

分 类 号：Q963[生物学—昆虫学]