大鼠睾丸AQP8和绿色荧光蛋白融合蛋白表达载体构建  

Reconstruction of aquaporin 8 and green fluorescent protein fusion recombinant vector

作　　者：梁爽[1] 于振香[2] 赵丹[1] 杨柯[1] 赵雪俭[1] 杨宝学[3] 

机构地区：[1]吉林大学基础医学院生殖病理生理研究室,吉林长春130021 [2]吉林大学第一医院呼吸内科,吉林长春130021 [3]美国加利福尼亚大学医学院,美国旧金山ca94122

出　　处：《吉林大学学报（医学版）》2006年第2期218-220,共3页Journal of Jilin University：Medicine Edition

基　　金：国家自然科学基金资助课题课题(30370572)

摘　　要：目的：克隆Wistar大鼠睾丸水通道蛋白（AQP8）cDNA，构建增强型绿色荧光蛋白（EGFP）与大鼠AQP8的融合蛋白真核细胞表达载体。方法：用RT-PCR钓取AQP8 cDNA编码区全长序列，测序鉴定，将AQP8 cDNA亚克隆于pEGFP-C1基因的下游。结果：①AQP8 cDNA测序结果与登录在GenBank上的大鼠AQP8基因序列（GenBank登录号：NM_019158）高度同源，但PCR产物的序列分析结果中发现4个碱基序列与已发表的AQP8序列不同：NM_019158的第135～137位碱基分别是C、A和G，而测序结果缺乏这3个碱基，NM_019158的第311位为A，而测序结果为G；②酶切鉴定在表达载体pEGFP—C1中AQP8cDNA融合于EGFP基因的下游。结论：pEGFP-C1-AQP8融合蛋白的表达载体构建成功。Objective To clone aquaporin 8 （AQP8） cDNA of testis in Wistar rats and construct the eucaryotic expression vector of enhance green fluorescent protein （EGFP） and AQP8 fusion protein. Methods The full sequence of AQP8 was fished by RT-PCR and sequenced, then it was recombined in the downstream of the pEGFP- C1 gene. Results The sequence of AQP8 cDNA of Wistar rat was homology with NM _ 019158 （access number logged in GenBank）, but there were 4 different bases： P135-137 of NM_ 019158 partly were C, A and G, but the sequencing result was absence; P311 of NM _ 019158 was A, but the sequencing result was G. The recombinant pEGFP-C1-AQP8 was identified by enzyme digestion. Conclusion The recombinant pEGFP-C1-AQP8 is constructed successfully.

关 键 词：水通道 融合蛋白 绿色荧光蛋白 睾丸 大鼠 

分 类 号：Q78[生物学—分子生物学] R-332[医药卫生]