视网膜发育基因Six3表达载体的构建  

Construction of expression vector of development gene Six3 in retina

作　　者：孙雪荣[1,2] 葛坚[1] 章晓霜[2] 邓飞[1] 胡惠玲[1] 

机构地区：[1]中山大学中山眼科中心眼科学国家重点实验室,广东广州510000 [2]湛江师范学院体育科学学院,广东湛江524000

出　　处：《海南医学院学报》2011年第3期289-291,共3页Journal of Hainan Medical University

基　　金：国家自然科学基金(30973266;30672275);眼科学国家重点实验室青年基金(QN-01);湛江师范学院博士专项基金(ZL1014)~~

摘　　要：目的:Six3在视网膜发育中起着关键性的调节作用,然而商品化的Six3表达载体尚鲜见,本研究拟以pBaBb-puro为骨架,构建Six3真核表达载体。方法:以新生乳鼠眼cDNA为模板,通过PCR扩增出Six3编码序列,经酶切后,用T4 DNA连接酶将其与pBaBb-puro进行连接,经过转化、筛选后,进行酶切和测序鉴定。结果:经RT-PCR扩增获得了含1002bp的Six3基因编码序列,经过酶切、连接、转化后,挑选12个菌落进行PCR检测,筛查到9个菌落含有重组质粒,对其中2个菌落进一步行酶切和测序鉴定,结果与理论预期完全相符。结论:成功构建了pBaBb-puro-Six3真核表达载体,为进一步研究Six3的功能奠定了基础。Objective：To investigate the construction of eukaryotic expression vector of Six3 with pBaBb-puro as bone.Methods：cDNA of eyes in neonatal murine was selected as template,Six3 encoding sequence was amplified by PCR.After enzyme digestion,this sequence was linked with pBaBb-puro by T4 DNA ligase.After transformation and screening,enzyme digestion and sequencing were performed.Results：The100 2 bp encoding sequence of Six3 gene was obtained.Twelve colonies were selected for detection after enzyme digestion,linkage and transformation.It revealed 9 colonies with recombinant plasmid.And 2 colonies were selected for further enzyme digestion and sequencing.It showed the expected result.Conclusions：Eukaryotic expression vector pBaBb-puro-Six3 is successfully established.It provides foundation for further research on Six3.

关 键 词：Six3 视网膜 pBaBb-puro 表达载体 鼠 

分 类 号：R774.1[医药卫生—眼科]