摘要
目的:获得由插入序列ISS1构建的德式乳杆菌保加利亚亚种胸腺嘧啶合成酶thy A基因缺陷型菌株。方法:利用两端带有ISS1的载体p Ghost9:ISS1,经复制型转座构建突变体,以50μg/m L脱氧胸苷、梯度浓度三甲氧苄胺嘧啶(TMP)连续富集传代培养,以添加终质量浓度50μg/m L脱氧胸苷的SA平板以及未添加脱氧胸苷的SA平板筛选thy A基因缺陷型菌株。结果:获得生物学性状稳定,回复突变率低的thy A基因插入突变株。
Objective： Using the insertion sequence ISS1 construction mutants, in order to isolate the Lactobacillus delbrueckii subsp. bulgaricus thy A-null strains. Methods： After ISS1 replicative transposition, the chromosome contains duplicated ISS1 sequences flanking p Ghost, Lactobacillus delbrueckii subsp. bulgaricus is efficient mutagenesis. The mutants were cultivated on MRS supplement with deoxythymidine（50 μg·m L-1） and TMP（1 to 500 μg·m L-1, depending on the strain）. After about 33 generations of cultivation, each culture broth was spread on MRS agar plates containing deoxythymidine. Those single-colonies isolates of each strain were examined by growth on SA agar with and without added deoxythymidine. Results： Achieving the thy A-null strains with the insetion of ISS1 and the mutants are stable.
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2015年第11期23-28,共6页
Journal of Chinese Institute Of Food Science and Technology
基金
国家高技术研究发展计划(863计划)项目(No.2006AA10Z317)
国家自然科学基金项目(31301520)
河北省高等学校科学技术研究项目(QN2014065)
