Balb/c小鼠胚胎皮肤无瘢痕愈合相关基因cDNA文库构建  

The construction of scarless wound healing related gene cDNA library from Balb/c mouse embryonic skin

作　　者：王鹏[1] 黄勇[1] 林立新[1] 姜蕾[1] 

机构地区：[1]烟台毓璜顶医院,山东264000

出　　处：《医学动物防制》2014年第5期484-486,489,共4页Journal of Medical Pest Control

摘　　要：目的筛选、克隆Balb／c成年小鼠和小鼠胚胎皮肤的特异表达基因，构建成年小鼠和小鼠胚胎皮肤差异表达的cDNA文库。方法应用Trizol法抽提样本的总RNA，进一步获得mRNA，结合PCRcDNA合成法将合成的双链cD-NA产物与载体连接构建cDNA文库，并转染大肠杆菌进行文库扩增，菌液进行PCR扩增，筛选有插入片段cDNA克隆。结果成功地构建了Balb／c小鼠皮肤无瘢痕愈合相关基因差异表达的cDNA文库，从中挑取85个克隆进行菌液PCR分析，结果显示57个克隆得到300—9Kp插入片段。结论通过RT—PCR方法并合成双链cDNA是一种自微量总RNA中构建高特异性的差异表达cDNA文库的有效方法。cDNA文库的构建有助于筛选、克隆小鼠胚胎皮肤无瘢痕愈合相关的特异表达基因。Objective To screen and clone Balb / c mice and mouse embryonic adult skin specific genes, con-structed adult mice and mouse embryonic skin differentially expressed cDNA library. Methods The sample was extracted by Trizol total RNA, further access to mRNA, with PCRcDNA synthesis, the synthesized double - stranded cDNA product was linked to carriers to construct a cDNA library, and transfected into E. coli, for PCR amplification bacteria increasing screening a cDNA clone insert. Results Successfully constructed Balb /c mouse skin wound healing associated genes differentially expressed cDNA library, from which 85 clones were picked, bacteria PCR analysis showed that among 57 clones obtained 3 0 0 - 9Kp inserts. Conclusions It is an effective way of synthesized by RT - PCR and the double - stranded cDNA to a trace of total RNA from the build high specificity for differentially expressed cDNA library. The construction of cDNA library can help the expression of related genes of screening and cloning of mouse embryonic skin wound healing.

关 键 词：无瘢痕愈合 基因 CDNA文库 

分 类 号：R619.6[医药卫生—外科学]