恒河猴Fas基因的克隆

目的研究猴艾滋病发病机制中细胞凋亡的作用,克隆凋亡相关基因Ｆａｓ的ｃＤＮＡ序列。方法从恒河猴腹股沟淋巴结中提取总ＲＮＡ,根据人的Ｆａｓ ｃＤＮＡ序列设计上、下游引物,通过ＲＴ－ＰＣＲ扩增出目的ｃＤＮＡ片段,将这一片段克隆到ｐＧＦＭ－Ｔ Ｅａｓｙ载体中,筛选阳性克隆并进行序列测定。结果首次克隆了恒河猴Ｆａｓ基因,编码序列为１００５ ｂｐ,将其序列提交ＧｅｎＢａｎｋ,收录号为ＡＹ００７５７２。结论克隆的新序列与ＧｅｎＢａｎｋ已收录的人和食蟹猴的Ｆａｓ基因在编码序列的长度上稍有区别,人的编码序列为１００８ｂｐ,食蟹猴的为９９６ ｂｐ。

Cloning of Fas Gene in Rhesus Monkey

Objective To investigate the role of apoptosis in the pathogenesis of simian acquired immunodeficiency syndrome (SAIDS), we cloned the cDNA fragment of Fas gene in Rhesus monkey. Methods We extracted total RNA from inguinal lymph nodes and designed the upper and low primers according to the Fas gene of human. The fragment was obtained by RT-PCR, and then cloned into PGEM-T Easy vector Finally, the positive clone was sequenced. Results We first cloned the Fas gene of Rhesus monkey, the encoding sequence is 1 005 bp.The sequence was submitted to GenBank, accession number is AY007572. Conclusion The length of encoding sequence in Rhesus monkey Fas gene is different from the homologous genes of human and cynomolgus monkey which have already been submitted to GenBank.