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| 抗菌肽CM与haFGF改构体融合基因的构建与表达 | |
| 引用本文： | 解佳森,肖业臣,秦玉侠,王会岩,张继威,李校垄.抗菌肽CM与haFGF改构体融合基因的构建与表达[J].吉林农业大学学报,2009,31(4). |
| 作者姓名： | 解佳森 肖业臣 秦玉侠 王会岩 张继威 李校垄 |
| 作者单位： | 1. 吉林农业大学生命科学学院,长春,130118 2. 生物反应器与药物开发教育部工程研究中心,长春,130118 3. 吉林农业大学中药材学院,长春,130118 |
| 基金项目： | 吉林省科技发展计划项目，长春市净月科技三项费用项目，吉林农业大学博士启动基金项目 |
| 摘 要： | 构建重组改构体aFGF28-154和抗菌肽CM基因的融合基因,使其在大肠杆菌系统中融合表达,以期获得具有靶向FGFR高表达肿瘤细胞的抗肿瘤融合蛋白.利用PCR引物延伸的方法拼接得到带有G4S铰链的天蚕素-蜂毒素杂合肽CM与aFGF28-154的融合基因,插入大肠杆菌表达载体pET20b中,构建表达质粒pET-CM-aFGF28-154,并转化至大肠杆菌BL21(DE3),氨苄青霉素抗性筛选重组转化子.IPTG诱导4 h后,菌液经超声破碎,以包涵体形式表达的融合蛋白约占菌体总蛋白的18%.将包涵体溶解并通过差速离心除去部分杂蛋白,透析复性并用Heparin Sepharose CL-6B亲和层析,目的蛋白得到初步纯化,为进一步研究蛋白的生物学活性奠定了基础. |
| 关 键 词： | 天蚕素-蜂毒素杂合肽CM 人酸性成纤维细胞生长因子改构体aFGF28-154 融合基因 |
Construction and Expression of Fusion Gene of Antibacterial Peptide CM and Modified haFGF |
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| XIE Jia-sen,XIAO Ye-chen,QIN Yu-xia,WANG Hui-yan,ZHANG Ji-wei,LI Xiao-kun.Construction and Expression of Fusion Gene of Antibacterial Peptide CM and Modified haFGF[J].Journal of Jilin Agricultural University,2009,31(4). | |
| Authors: | XIE Jia-sen XIAO Ye-chen QIN Yu-xia WANG Hui-yan ZHANG Ji-wei LI Xiao-kun |
| Institution: | XIE Jia-sen1,XIAO Ye-chen2,QIN Yu-xia2,WANG Hui-yan2,ZHANG Ji-wei3,LI Xiao-kun21.College of Life Science,Jilin Agricultural University,Changchun 130118,China,2.Ministry of Education,Engineering Research Center of Bioreactor , Pharmaceutical Development,3.College of Chinese Medicinal Materials |
| Abstract: | The fusion gene of modified aFGF28-154 and antibacterial peptide CM was recombinated and expressed in the E.coli system so as to obtain fusion protein targeting tumour cells which have high expression of FGFR.Firstly,the gene of Cecropin A1-7,melittin5-12and aFGF28-154 was connected with G4S hinge by the method of PCR,inserted into expression vector pET20b to obtain expression plasmid pET-CM-aFGF28-154,and then transformed into BL21(DE3).Transformant was selected by ampicillin resistance.Four hours after be... |
| Keywords: | CM aFGF28-154 fusion gene |
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