大鼠Fas配体基因的克隆  

THE CLONING OF FAS LIGAND GENE IN THE RAT

作　　者：孙成三[1] 段德义[2] 于凤山[2] 于恩华[1] 徐群渊[2] 

机构地区：[1]北京医科大学解剖学系,北京100083 [2]首都医科大学神经科学研究所,北京100054

出　　处：《解剖学报》2000年第2期177-179,共3页Acta Anatomica Sinica

摘　　要：目的　研究 Fas配体 (Fas ligand,Fas L )在移植免疫方面的作用 ,克隆其编码的全部 c DNA序列 ,并添加 Kozak序列 ,以便在真核细胞高效表达大鼠 Fas L。　方法　从大鼠的睾丸组织中提取总 RNA,设计上、下游引物以 RT- PCR的方法扩增出一 931bp的 DNA片段 ,将这一片段重组于 p BK - RSV载体中 ,酶切鉴定插入片段正确后进行全序列分析。　结果　证实该研究所克隆的 c DNA是编码正确的大鼠 Fas L基因 ,与发表于 Gene Bank上的序列相比 ,完全正确。已构建成用于真核表达的重组质粒 p BK- RSV/r Fas L。　结论　克隆到了编码正确的大鼠Fas L的 c DNA全序列 ,对移植免疫和帕金森病基因治疗的基础研究及临床应用具有重要意义。Objective\ The cDNA fragment of rat Fas ligand with additional Kozak sequence was cloned in order to investigate the roles of FasL in gene therapy for transplant\|immune and its expression in eukaryotic cells.Methods\ 100mg tissue of rat testis was used to extract total RNA, a 931bp positive DNA band was obtained by reverse transcription PCR(RT\|PCR) method. The fragment was then cloned into the pBK\|RSV vector and the insert was sequenced after analysis of the recombinant pBK\|RSY plasmids by restriction enzymes. Results\ The DNA fragment of 931bp with additional Kozak sequence was amplified from reverse transcription mixture, and it proved to be correct compared with the sequence deposited in GeneBank(U03470). The recombinant eukaryotic expression plasmid(pBK\|RSV/rFasL) was then constructed. Conclusions\ The full\|length cDNA encoding FasL with additional Kozak sequence was harvested and can be expressed in eukaryotic cells.

关 键 词：基因克隆 DNA FAS配体 大鼠 震颤性麻痹 

分 类 号：R742.5[医药卫生—神经病学与精神病学] R394[医药卫生—临床医学]