摘要
为了研究人碱性成纤维细胞生长因子在治疗神经系统疾病中的作用 ,克隆其 c DN A序列 ,并添加 K ozak序列 ,以用于真核细胞表达。本实验提取我国自建的脑胶质瘤细胞系 BT32 5总 RN A,设计上、下游引物用逆转录 PCR法扩增 c DNA片段 ,然后重组于 p GEM- 3zf( + )载体 ,酶切鉴定插入片段正确后测序。结果 :RT- PCR扩增到 4 73bp的带有 K ozak序列的 c DN A片段。显示 :克隆到碱性成纤维细胞生长因子 c DN A片段 。
The cDNA fragment of human bFGF with additional Kozak sequence was cloned in order to investigate the roles of bFGF for treatment of diseases in the nervous system and to get its expression in eukaryotic cells. Total cellular RNA of glioma BT325, a cell line established from a Chinese patient, was isolated and reverse transcription PCR amplification of the cDNA fragments was performed. The fragment was then cloned into pGEM 3zf(+) vector and the inserts were sequenced after analysis of the recombinant pGEM 3zf(+) plasmids by restriction enzymes. The DNA fragment of 473 bp with additional Kozak sequence was amplified from reverse transcription mixture. It is concluded that full length of cDNA of human bFGF with additional Kozak sequence was cloned and can be expressed in eukaryotic cells.
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2000年第2期131-134,共4页
Chinese Journal of Neuroanatomy
