FMS样酪氨激酶3受体配基基因的克隆和序列分析

Molecular cloning and sequencing of human c fms-like tyrosine kinase 3 receptor ligand gene

目的 获得人FLcDNA膜外序列 ,以构建表达人FMS样酪氨激酶 3受体配基 (FL)蛋白的重组体 ,更深入地探讨FL的生物学功能。方法 通过设计和合成引物 ,提取胎肝细胞总RNA进行逆转录反应 ,经PCR扩增目的cDNA片段 ,并将其重组至PUC 18T载体中 ,测定其DNA序列。结果 从胎肝细胞总RNA中扩增得到 5 46bp片段 ,序列测定证实该片段为包括 2 5个氨基酸残基组成的信号肽的胞外序列。

Objective To obtain the cDNA sequence encoding extracellular human c fms like tyrosine kinase 3 receptor ligand(FL) gene with multiple functions. Methods The primers are designed based on the known FL cDNA sequence. Total RNA was isolated from fetal liver cells, and RT PCR was performed. The fragment was recombined into PUC 18 T vector, and further sequenced by automatic sequence analyzer. Results The 546bp DNA fragment was amplified by RT PCR method from liver cells and its sequence is identical to the published sequence encoding human FL. Conclusion cDNA has been cloned successfully.