PCR克隆高GC含量牛Fcγ2R cDNA

Cloning of Bovine GC-rich Fcγ2R cDNA by PCR

根据已发表的牛Fcγ2RcDNA序列设计1对引物,采用改进的RT -PCR技术从牛外周血淋巴细胞总RNA中扩增出了GC含量为6 4%的牛Fcγ2RcDNA分子,克隆片段全长795个核苷酸,含有一个完整的开放阅读框,编码2 6 5个氨基酸,和已报道的序列相比存在3个碱基突变。

The cDNA of bovine Fcγ2R with a GC-content 64 percent was cloned by improved PCR using a pair of primers based on previously cloned boFcγ2R.The cloned boFcγ2R cDNA was 795 nucleotides long and deduced 265 amino acids. When it was compared with the published sequence, 3 mutant sites was found. The improved PCR method can be widely available for amplification of a GC-rich template.