非天然氨基酸插入法构建小鼠突变rankl蛋白的克隆、原核表达及抗血清制备

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1、分类号R681UDC617.3密级公开非天然氨基酸插入法构建小鼠突变RANKL蛋白的克隆、原核表达及抗血清制备李锋培养类别全日制学位类型学术学位一级学科(专业类)临床医学二级学科(专业)外科学（骨外）研究方向骨质疏松的防治及其机制研究指导教师陶惠人教授（主任医师）培养单位西京医院骨科二O一五年五月第四军医大学硕士学位论文目录缩略语表·······························································································1中文摘要·····

2、··························································································3英文摘要·······························································································5前言···············································································

3、················8文献回顾······························································································10正文······························································································19实验一小鼠RANKL基因的克隆··········································

4、················191实验材料及主要仪器·······································································191.1实验材料················································································191.2主要试剂···········································································

5、·····201.3主要试剂的配制·······································································201.4主要仪器················································································202方法··························································································

6、··212.1骨髓细胞总RNA提取·······························································212.2反转录合成cDNA·····································································212.3mRANKL胞外段基因的扩增························································222.4pET28a-mRANKL重组表达载体的构建·······

7、···································232.5pET28a-mRANKL重组表达载体的转化··········································232.6菌液PCR鉴定·········································································242.7重组表达质粒的酶切鉴定···························································242342.8

8、pET28a-pNO2PhemRANKL重组表达载体的构建···························243结果···································