大菱鲆β-actin基因的初步研究  

The Cloning and Analysis of the β-actin Gene in Turbot

作　　者：王家庆[1] 侯林[1] 李晓燕[1] 赵欣涛[1] 张瑞锋[1] 姜丽娟[1] 孙文静[1] 安家璐[1] 

机构地区：[1]辽宁师范大学生命科学学院,辽宁大连116029

出　　处：《水产科学》2007年第4期214-217,共4页Fisheries Science

摘　　要：脊椎动物中不同物种的β-actin基因氨基酸之间的序列同源性超过98%。根据GenBank中登录的大菱鲆β-actin基因mRNA部分序列,与川鲽该基因全编码区DNA序列比对,依据比对结果分别在大菱鲆β-actin基因相邻外显子上设计1对引物,分别以RNA反转录产物和DNA为模板通过PCR方法进行克隆分析。对不同退火温度和循环数条件下的该基因RT-PCR产物进行了对比。针对RNA提取过程中的经过DNase处理但很难除净的痕量DNA污染,在琼脂糖凝胶电泳中很难被检测到的问题,初步探讨了利用上述2种模板对该基因扩增片断大小的不同来检出痕量DNA污染的可行性。Amino acid identities of β-actin gene are more than 98% among different vertebrates. According to the partial mRNA sequences of turbot （ Scophthatmus maximus） GenBank and the comparison with the same gene complete coding region of European flounder （ Platichthsys fiesus） in reverse transcript of mRNA and DNA templates using DNA sequence, cloning and analysis were conducted a pair of primers designed at adjacent exons. The RT-PCR products were compared under different annealing temperatures and cycles. The issues focussed mainly on the tracing DNA contamination which is difficult to eliminate in the process of RNA extraction and detection in agarose gel electrophoresis, so the feasibility of the detection of tracing DNA contamination according to the different sizes of β-actin PCR products between the two templates were preliminarily discussed.

关 键 词：大菱鲆 Β-ACTIN 克隆分析 痕量DNA污染 

分 类 号：Q953[生物学—动物学]