三种猕猴属动物Fas基因的克隆

Cloning of Fas Gene in Three Macaca Species

从腹股沟淋巴结或血液中提取总RNA ,根据人的Fas基因的cDNA序列设计上、下游引物 ,通过RT -PCR扩增出三种猕猴属动物Fas基因的cDNA片段 ,将片段克隆到 pGEM -TEasy载体中 ,筛选阳性克隆并进行序列测定。恒河猴Fas基因编码序列为 10 0 5bp ,GenBank收录号为AY0 0 75 72 ;熊猴的编码序列为 996bp ,GenBank收录号为AF32 6 2 0 8;短尾猴的编码序列为 933bp ,GenBank收录号为AF332 35 7。对五种已知的灵长类动物的Fas基因进行序列比较 ,结果表明Fas基因的保守区域具有高度保守性 。

Total RNA were extracted from inguinal lymph nodes or blood.The Fas cDNA fragments of three macaca species were obtained by RT-PCR using the upper and low primers according to the Fas gene of human,and then cloned into pGEM-T Easy vector.The positive clones were sequenced.For the first time we cloned the Fas genes of Macaca mulatta,Macaca assamensis and Macaca arctoides ,the encoding sequences are 1005bp, 996bp and 933bp, respectively.Finally we compared the sequences of Fas genes among five primate species that have already been submitted to GenBank.The results confirmed the conservation of regions in two ends of Fas genes,and the high variability of a low complexity region in middle of every Fas gene.