特异性检测胰腺组织表达Cre重组酶转基因小鼠的方法

利用组织特异性分子标志物启动子调控Cre重组酶,研制了6种在不同组织中特异性表达Cre重组酶的转基因小鼠.这些转基因小鼠的基因型鉴定均使用设计在Cre基因编码区的通用引物.为了特异性检测胰腺组织表达Cre重组酶的转基因小鼠,在大鼠胰岛素RIP启动子上和Cre基因上设计1对引物进行PCR扩增,并通过凝胶电泳进行分析.PCR结果显示,设计在Cre基因上的通用引物可以从6种不同组织特异性Cre重组酶转基因小鼠基因组DNA中扩增获得480 bp产物;利用本研究设计的特异性引物可以从胰腺组织表达Cre重组酶转基因小鼠基因组DNA中扩增200 bp的目的条带.这一结果表明,利用特异性引物进行PCR反应,可有效地将胰腺组织表达Cre重组酶转基因小鼠与其他多种组织的Cm重组酶转基因小鼠鉴别开来.

Identification of the Pancreas Specific Cre Transgenic Mice Using PCR Analysis

We have generated six kinds of Cre transgenic mice in which the Cre recombinase driven by different tissue specific molecular marker promoter is expressed specifically in different tissues. The genotyping of these transgenic mice were performed using a pair of common primers designed in the coding region of Cre gene. In order to identify the pancreas specific transgenic mice which pancreas specific Cre recombinase expression, a pair of primers in the rat insulin promoter(RIP) and Cre gene were designed. The result of PCR analysis showed that a 480 bp fragment could be amplified from the genomic DNAs of all six tissue specific Cre recombinase transgenic mice using the common primers, while a 200 bp fragment could be obtained only from the genomic DNAs of the pancreas specific Cre transgenic mouse using the specific primers. The results indicated that we could distinguish the pancreas specific Cre transgenic mice from other tissue specific Cre transgenic mice by PCR analysis using the specific primers designed in this study.