正常成人肝组织中BMP-9读码框架的克隆  

CLONING OF BMP-9 OPEN READING FRAME IN ADULT LIVER TISSUE

作　　者：杨堃[1] 刘相萍[1] 隋爱华[1] 吴力群[2] 吕振华[1] 

机构地区：[1]青岛大学医学院附属医院分子生物学中心实验室,山东青岛266003 [2]青岛大学医学院附属医院肝胆外科,山东青岛266003

出　　处：《青岛大学医学院学报》2007年第6期520-522,共3页Acta Academiae Medicinae Qingdao Universitatis

基　　金：山东省卫生厅青年基金资助项目(2005JZ004)

摘　　要：目的获得骨形态发生蛋白(BMP-9)基因并进行基因读码框架的克隆。方法自正常成人肝组织提取总RNA,一步法RT-PCR扩增BMP-9,扩增产物经过胶回收及酶切后,克隆入高效真核表达质粒pcDNA4/HisMax,构建重组表达载体。结果RT-PCR扩增获得约1.3kb大小的产物,经限制性酶切分析和DNA测序,证实为人BMP-9基因的完整开放读码框架。重组载体表达读码序列正确。结论正常成人肝组织总RNA经一步法RT-PCR,能够对BMP-9基因的开放读码框架进行完整有效的筛选。BMP-9开放读码框架的完整克隆为其在骨、内分泌和神经系统等多能作用的转基因实验研究提供了基础。Objective To clone BMP-9 gene in adult liver tissue and construct recombinant eukaryotic expression vectors. Methods Total RNA was extracted from normal liver tissue of an adult, then open reading frame of BMP-9 was cloned by one-step RT-PCR. The PCR products （1.4 kb） were inserted into eukaryotic expression vector, pcDNA4/HisMax, to construct recombinant vector. Results According to restrictive analysis and DNA sequencing of recombinant vector, the open reading frame of BMP 9 was identified and thc insert in MCS was correct. Conclusion The ORF of BMP-9 gene can be screened effectively from total RNA of normal liver tissue by one-step RT-PCR. The complete clone of ORF of BMP 9 has provided the basis for multi-function gene transfer experiments in the fields of orthopedics, endocrinology and central nervous system.

关 键 词：骨形态发生蛋白质类 重组真核表达载体 逆转录聚合酶链反应 

分 类 号：R346[医药卫生—基础医学]