摘要
目的 构建人AQP1(hAQP1)基因的重组腺病毒载体。方法 采用DNA重组与细菌内同源重组技术 ,构建含hAQP1的复制缺陷型重组腺病毒载体 (pAdEasy 1 hAQP1) ,并观察pAdEasy 1 hAQP1在ECV 3 0 4中的感染情况。PCR方法鉴定重组腺病毒载体 ,利用绿色荧光蛋白GFP检测病毒滴度和感染效率。结果 PCR及限制性内切酶酶切鉴定证明pAdEasy 1 hAQP1构建成功。
Objective To construct the recombinant adenovirus of human aquaporin 1 (AQP1) gene. Methods The replication deficient recombinant adenovirus encoding hAQP 1(pAdEasy 1/ hAQP1) was constructed by the method of DNA recombination and homogenous recombination in bacteria. The ECV 304 cells were transfected by the recombinant adenovirus. The target gene was identified by polymerase chain reaction (PCR). The titer and the infection rate were determined using the green fluorescent protein (GFP) expression in the shuttle plasmid. Results Restriction endonuclease and PCR analyses confirmed that the recombinant adenovirus was successfully constructed. The infection rate was high. Conclusion The recombinant adenovirus pAdEasy 1/ hAQP1 has been successfully constructed by the method of homogenous recombination in bacteria.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2004年第6期530-532,共3页
Journal of Third Military Medical University
基金
重庆市科技项目计划资助项目 ( 2 0 0 30 6 0 8)~~
关键词
腺病毒
AQP1
同源重组
adenovirus
AQP1
homogenous recombination
