用PCR直接筛选和鉴定虎纹捕鸟蛛毒素I的重组阳性克隆  

Screen and identification for recombinant clones of HWTX-I by polymerase chain reaction

作　　者：何宁佳[1] 聂东宋[1] 梁宋平[1] 

机构地区：[1]湖南师范大学生命科学院,长沙410081

出　　处：《生物技术通讯》2001年第3期211-212,共2页Letters in Biotechnology

摘　　要：以合成的两段插入序列为上、下游引物用PCR法直接筛选插入有虎纹捕鸟蛛毒素I(HWTX I)cDNA的重组阳性克隆。并用PCR法快速鉴定重组体中插入片段的正、反连接方向 ,扩增用引物是以位于克隆位点上游的一段载体序列为上游引物 ,以插入序列为下游引物。对 10 0个单克隆进行了上述两次PCR筛选鉴定 ,选取 2个有靶片段插入并且为正向连接的重组子进行测序 。Using two digomers of insert fragment as upstream and downstream primers separately,a rapid PCR method was developed to screen the Huwentoxin-I(HWTX-I) cDNA recombinant clones. PCR technique was also designed to identify the orietation of inserts in the recombinants. In this PCR reaction, the sequence near the cloning site of vector was selected as upstream primer, and one oligomer of the insert as downstream primer separately. One hundred clones were screened and identified by above two PCR reactions and two positive clones were further checked by DNA sequencing. The sequencing results confirmed the positive clones and theirs correct directions.

关 键 词：PCR 虎纹捕鸟蛛毒素Ⅰ 重组体 鉴定 筛选 

分 类 号：R996.3[医药卫生—毒理学] Q78[医药卫生—药学]