假单胞杆菌BS1转座突变及高产生物表面活性剂突变株初步筛选

生物技术通报 ›› 2014, Vol. 0 ›› Issue (5): 179-183.

假单胞杆菌BS1转座突变及高产生物表面活性剂突变株初步筛选

侯巨梅¹ 韩巍¹ 国莉玲² 刘铜² 王彦杰³

(1. 黑龙江八一农垦大学农业部农产品质量安全监督检验测试中心，大庆 163319；2. 黑龙江八一农垦大学农学院植物病理与应用微生物研究所，大庆 163319；3. 黑龙江八一农垦大学生命学院，大庆 163319)

收稿日期:2014-02-26 出版日期:2014-05-23 发布日期:2014-05-24
作者简介:侯巨梅，女，硕士，研究方向：微生物学；E-mail：amyliutong@163.com
基金资助:
黑龙江教育厅科学技术研究项目(12511326)

Transposon Mutagenesis of Pseudomonas sp. BS1 and Preliminary Screening of High-producing Biosurfactant Mutant

Hou Jumei¹ Han Wei¹ Guo Liling² Liu Tong² Wang Yanjie³

(1. Supervision ＆Testing Center for Agricultural Product Quality，Ministry of Agriculture，Heilongjiang Bayi Agricultural University，Daqing 163319；2. Institute of Plant Pathology and Applied Microbiology，Agriculture School，Heilongjiang Bayi Agricultural University，Daqing 163319；3. College of Life Science and Technology，Heilongjiang Bayi Agricultural University，Daqing 163319)

Received:2014-02-26 Published:2014-05-23 Online:2014-05-24

摘要/Abstract

摘要： 利用含转座子Tn917的温敏性质粒pTV1-OK转化假单胞杆菌BS1原生质体，成功获得3个稳定的转化子；通过Tn917诱导转座突变，产生大量突变体，构建了转座突变体库，采用特异引物对随机挑取的5株突变体进行PCR扩增，获得与预期大小一致片段，表明突变体基因组中有Tn917插入；通过对随机挑取24株突变体乳化(E24)性能测定，发现有一株突变体E24值达到67%，明显高于野生菌株。结果表明转座突变是假单胞杆菌BS1获得高产生物表面活性剂菌株的一种有效手段。

关键词: 原生质体, 转座, 乳化性能, 生物表面活性剂

Abstract: The protoplast of Pseudomonas sp. BS1 was transformed by temperature-sensitive plasmid pTV1-OK that harbor Tn917. 3 stable transformants were obtained from the LB plates with Kan and Erm. Lots of mutants were obtained by transposon mutagenesis of Tn917, sequencely a mutant library was constructed. A predicted fragment was amplified by PCR from 5 randomly selected mutants, which showed that Tn917 successful inserted into the genome of Pseudomonas sp. BS1. E24 of a mutant from 24 selected randomly reach up 67% by the measurement of the emulsification index, significantly higher than wild-type strain. These results indicated that the transposon mutagenesis is an effective method for obtaining high-producing biosurfactant.

Key words: Protoplast, Transposon, Emulsification, Biosurfactant

侯巨梅, 韩巍, 国莉玲, 刘铜, 王彦杰. 假单胞杆菌BS1转座突变及高产生物表面活性剂突变株初步筛选[J]. 生物技术通报, 2014, 0(5): 179-183.

Hou Jumei, Han Wei, Guo Liling, Liu Tong, Wang Yanjie. Transposon Mutagenesis of Pseudomonas sp. BS1 and Preliminary Screening of High-producing Biosurfactant Mutant[J]. Biotechnology Bulletin, 2014, 0(5): 179-183.

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