摘要
通过ＰＣＲ二步法，构建了ｍＣＳＦ－１Ｒ激酶负性突变子，以及野生型和突变型ＣＳＦ－１Ｒ的送病毒表达载体ｐＣＥＮ／ＭＰＳＶ。进行了１２５Ｉ－ＣＳＦ－１受体结合分析：检测了激酶负性突变子，删除了ＣＳＦ－１ＲＣ－末端氨基酸９２５以后部分的突变体（ＣＴＲＵＮＣ９２５）和删除了激酶插入区的突变体（△ＫＩ）在３２Ｄ髓细胞表达。表达水平可达１～２×１０４受体／细胞。初步测定了通过ＣＳＦ－１Ｒ所介导的促细胞分裂效应。
Author constructed CSF-1R kinase minus mutant(K614M)using PCR 2 steps and reverse exprea vector pCEN/MPSV for wild-tye and mutants of CSF-1R. Their expression in murine 32D myeloid cells included K614M and 2 truncation mutants, CTRUNC 925 (absent C-terminus) and KI (absent KI) were identified by receptor bind assay with 125I-CSF-1. Usually, expression level reached 1-2×104 receptors/cell.The effect of mitogenesis mediated by CSF-1R was analyzed.
出处
《高技术通讯》
CAS
CSCD
1996年第6期52-56,共5页
Chinese High Technology Letters
关键词
突变体
细胞表达
激酶突变子
CSF-1R,Mutant,Receptor,Cellular expression
