应用计算机软件辅助重组hbFGF基因的筛选  

Selecting the recombinant hbFGF gene under the help of computer software

作　　者：黄更生[1] 孙奋勇[2] 卜丹 

机构地区：[1]广东教育学院生物系,广州510303 [2]暨南大学生物工程研究所,广州510632 [3]广州华侨医院,广州510630

出　　处：《现代生物医学进展》2006年第8期19-21,共3页Progress in Modern Biomedicine

摘　　要：目的:尝试通过计算机的辅助筛选基因重组组合,期望能减少人工筛选的工作量并得到高表达的蛋白。方法:从影响表达的两个因素即翻译起始区TIR的二级结构自由能和翻译起始区的密码子偏好性问题研究入手,对部分核苷酸进行了突变,利用RNA二级结构预测软件DNASISv2.5对hbFGF的TIR区起始密码子开始前35个核苷酸进行了分析,筛选出预期表达量高的组合并在实验中加以验证。结果:从32种突变方式中筛选出10条自由能绝对值最低的序列做引物,克隆至表达载体上,得到了两株高表达菌株。结论:利用计算机辅助设计可以优化和筛选实验结果,提高工作效率,降低具体实验的工作量。Background＆Objective： We try to use softwares to cut down our workload and to have a higher protein expression. Methods： Tire free energy of the Translation Initiation Region（TIR） and the Codons preference of E. cull were started with. During the cloning, there are two principles of increasing the region, which is mainly decided by the content of G ＋ C in TIR. We adjusted the content of G ＋ C in the 35 aminophenol near the N terminal of hbFGF, then worked out the △GO with software DNASISv2.5 and screened the clones with higher expression. Results; Calculating by the software, we got 10 sequence from all possible combination with lower △G0 and cloned them into expression cell BL21 （DE3）. Two of the candidates showed high express potential. Conculusion： Utilizing the auxiliary design of computer software can not only optimize and select experimental results, but also reduce the workload of specific experiment and raise working efficiency.

关 键 词：计算机 DNAsia2.5 HBFGF 自由能 

分 类 号：Q786[生物学—分子生物学]