| 黄更生1 孙奋勇2 卜 丹3.应用计算机软件辅助重组hbFGF 基因的筛选[J].现代生物医学进展英文版,2006,6(8):19-21. |
| 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 |
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| DOI： |
| 中文关键词: 计算机 DNAsis2 .5 hbFGF 自由能 |
| 英文关键词: Computer DNAsis 2.5 hbFGF Free energy |
| 基金项目: |
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| 中文摘要: |
| 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。 |
| 英文摘要: |
| Background&Objective :We try to use softwares to cut down our workload and to have a higher protein expression.Methods:
The free energy of the Translation Initiation Region(TIR)and the Codons preference of E.coli 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 ■G0 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 . |
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