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中国精品科技期刊2020
张广峰, 张楠笛, 赵婷婷, 许琴, 谭霄, 向文良. 一株食窦魏斯氏菌的分离鉴定及其质粒的序列分析[J]. 食品工业科技, 2018, 39(8): 100-105. DOI: 10.13386/j.issn1002-0306.2018.08.019
引用本文: 张广峰, 张楠笛, 赵婷婷, 许琴, 谭霄, 向文良. 一株食窦魏斯氏菌的分离鉴定及其质粒的序列分析[J]. 食品工业科技, 2018, 39(8): 100-105. DOI: 10.13386/j.issn1002-0306.2018.08.019
ZHANG Guang-feng, ZHANG Nan-di, ZHAO Ting-ting, XU Qin, TAN Xiao, XIANG Wen-liang. Isolation and identification of Weissella cibaria and sequence analysis of its plasmid[J]. Science and Technology of Food Industry, 2018, 39(8): 100-105. DOI: 10.13386/j.issn1002-0306.2018.08.019
Citation: ZHANG Guang-feng, ZHANG Nan-di, ZHAO Ting-ting, XU Qin, TAN Xiao, XIANG Wen-liang. Isolation and identification of Weissella cibaria and sequence analysis of its plasmid[J]. Science and Technology of Food Industry, 2018, 39(8): 100-105. DOI: 10.13386/j.issn1002-0306.2018.08.019

一株食窦魏斯氏菌的分离鉴定及其质粒的序列分析

Isolation and identification of Weissella cibaria and sequence analysis of its plasmid

  • 摘要: 目的:探究四川泡菜中含质粒的乳酸菌,并对质粒进行序列和功能分析。方法:采用平板涂布法和碱裂解法分离含质粒乳酸菌,通过菌落形态、生理生化和16S rDNA序列分析法对其进行鉴定。利用单酶酶切质粒,并在测序质粒片段的基础上设计引物扩增质粒的剩余片段,对拼接成功的质粒进行序列分析。结果:从四川泡菜中分离出一株含质粒的食窦魏斯氏菌W5(Weissella cibaria)。该菌株中质粒pA010全长为11630 bp,GC含量为34.6%,编码了4个可识别功能蛋白,包括酪氨酸重组酶、复制蛋白、抗毒素PemI和毒素PemK。比较酪氨酸重组酶和复制蛋白,推断该质粒为θ型复制质粒。其中PemI和PemK构成的Ⅱ-TA系统对质粒的稳定有一定作用且可能提升食窦魏斯氏菌对周围环境的适应力。结论:食窦魏斯式菌W5是一株含质粒乳酸菌,其序列分析为构建耐受环境胁迫的载体提供了可能。

     

    Abstract: Objective:Lactic acid bacteria harbored plasmid was isolated from Sichuan pickles and the sequence and function of plasmid was detected. Methods:The lactic acid bacteria harbored plasmid was screened by traditional microbial separation and alkaline lysis and then the strain was identified by morphological and physiological characteristics and its 16S rDNA sequence. Plasmid was digested with single restriction enzymes and the target DNA fragments were purified and sequenced. Then the whole sequence was finally elucidated by primer design and PCR cloning. Results:A strain harbored plasmids was isolated from Sichuan pickles and identified as Weissella cibaria. A large plasmid designated as pA010 was isolated from Weissella cibaria W5 and was 11630 bp in length with GC content of 34.6%. Sequence analysis indicated that the plasmid pA010 contained four known proteins,including tyrosine recombinase,replicative protein,antoxin PemI and toxin PemK. Based on tyrosine recombinase and replication protein,pA010 may belong to a theta-type replication plasmid. In addition,PemI and PemK,an Ⅱ-TA system,may have a role in the stability of the plasmid and enhance the adaptability of bacteria to the surrounding environment. Conclusion:Weissella cibaria W5 harbored plasmid and the sequence analysis of its plasmid pA010 provided a probability for construction vectors benefited to environmental stress tolerance.

     

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