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发明公开
CN101457231A 菌影载体PMAL-E-SN的构建及其在大肠杆菌中的应用 无效 - 驳回

菌影载体PMAL-E-SN的构建及其在大肠杆菌中的应用
摘要：
本发明公开了一种菌影载体PMAL－E－SN的构建及其在大肠杆菌中的应用，以PhiX174噬菌体RFI质粒为模板进行PCR扩增，得到E基因；以金黄色葡萄球菌标准株ATCC25923基因组DNA为模板进行PCR扩增，得到SN基因；用十五个柔性氨基酸的Linker串连E基因和SN基因，得到E－SN基因；E－SN基因PCR扩增产物与pMD18－T载体连接，筛选阳性质粒pT－E－SN测序；用质粒载体PMAL－p2X和测序正确的重组质粒pT－E－SN构建菌影载体PMAL－E－SN。本发明的菌影载体PMAL－E－SN通过“tac”强启动子和malE翻译起始信号使E基因和SN基因获得高效表达，成功制备了大肠杆菌菌影，裂解效率可达99.99％；本发明引入次要杀伤基因SN，并联合E基因和SN基因连入高效表达载体PMAL－p2X，取得了良好的效果，可广泛应用于大肠杆菌菌影的制备。
摘要（英）：

The invention discloses a bacterial ghost vector PMAL-E-SN construction and application thereof in an escherichia coli comprising an E gene obtained by PCR amplification using a PhiX174 phage RFI plasmid as a template; an SN gene obtained by PCR amplification using a staphylococcus aureus standard strain ATCC25923 genome DNA as a template; an E-SN gene obtained by serial connecting the E gene and the SN gene by the Linker of the fifteen flexible amino acids; a positive plasmid pT-E-SN sequencing screened by coupling an E-SN gene PCR amplification outcome and a pMD18-T vector and the bacterial ghost vector PMAL-E-SN constructed by the recombinant plasmid pT-E-SN with right sequencing and the plasmid vector PMAL-p2X. The bacterial ghost vector PMAL-E-SN of the invention enables the E gene and the SN gene to obtain the high efficiency expression through a ''tac'' strong promoter and a malE start signal, successfully prepares the escherichia coli bacterial ghost and the degradation efficiency can reach 99.99 The invention induces a secondary killing gene SN, obtains good effect by combining the E gene and the SN gene to a high efficiency expression vector PMAL-p2X and is widely used in preparation of the escherichia coli bacterial ghost.

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