摘要
目的KIF18A蛋白是调节有丝分裂和影响肿瘤发生发展的重要蛋白，本研究旨在建立KIF18A蛋白昆虫杆状病毒表达系统，从而可在体外高效合成KIF18A蛋白。方法先用分子克隆方法构建转移载体，然后通过基因转座作用得到重组杆状病毒，最后将重组杆状病毒转入昆虫细胞Sf-9以高效合成KIF18A蛋白。结果经过DNA测序、倒置显微镜下细胞观测和WesternBlot检测验证分析，证实已成功建立了KIF18A蛋白昆虫杆状病毒表达系统。结论明确了KIF18A杆状病毒表达系统中转移载体构建和重组病毒转染等条件，建立了高效表达KIF18A的杆状病毒表达系统。
Objective KIF18A is a protein that has close relation with mitotic regulation and tumor development. This study aimed to establish KIF18A protein expression system in baculovirus, which may help to realize high efficient synthesis of KIF18A protein in vitro. Methods Transfer vector was constructed with molecular cloning method, and recombinant baculovirus were obtained through gene transposition. KIF18A protein was expressed by transforming recombinant baculovirus into infected insect Sf-9 cells to realize the synthesis efficiency. Results It was confirmed by DNA sequencing, microscopic observation and Western Blot that the KIF18A protein expression system in baculovirus was successfully established. Conclusions Conditions for transfer vector and recombinant virus transfection are defined, and high efficient KIF18A protein baculovirus expression system are successfully constructed.
出处
《国际生物医学工程杂志》
CAS
2015年第2期73-76,I0004,共5页
International Journal of Biomedical Engineering
基金
基金项目：国家自然科学基金资助项目（31271485）
国家自然科学基金青年基金资助项目（31301138）
2011年教育部“新世纪优秀人才支持计划”资助项目（NCET-11-1066）
天津市应用基础与前沿技术研究计划重点资助项目（12JC2DJC21400）
天津师范大学中青年教授学术创新推进计划资助项目（52XBl104）
天津师范大学博士基金资助项目（52XCl001）
