Sequence Analysis of an Endogenous Plasmid from <i>Enterococcus faecalis</i> and the Construction of Shuttle Vectors

YUAN Lin; ZENG Jing; GUO Jianjun; WEI Guowen

doi:10.13386/j.issn1002-0306.2021040175

Volume 42 Issue 23

Nov. 2021

Turn off MathJax

Article Contents

Abstract

References

Science and Technology of Food Industry > 2021 > 42(23): 141-149. > DOI: 10.13386/j.issn1002-0306.2021040175

YUAN Lin, ZENG Jing, GUO Jianjun, et al. Sequence Analysis of an Endogenous Plasmid from Enterococcus faecalis and the Construction of Shuttle Vectors[J]. Science and Technology of Food Industry, 2021, 42(23): 141−149. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2021040175.

Citation:

PDF (2534 KB)

Sequence Analysis of an Endogenous Plasmid from Enterococcus faecalis and the Construction of Shuttle Vectors

Institute of Microbiology, Jiangxi Academy of Sciences, Nanchang 330096, China

More Information

Received Date: April 25, 2021
Available Online: September 29, 2021

Graphical Abstract

Abstract

Abstract

The aim of this study was to determine and analyze the DNA sequence of the endogenous plasmid pXW from Enterococcus faecalis EXW27, and to construct an Escherichia coli/Enterococcus faecalis shuttle vector based on its minimal replicon. The endogenous plasmid pXW was isolated from E. faecalis EXW27 with good probiotic properties. The DNA sequence of pXW was determined and analyzed, and then the replicon of plasmid pXW was used to construct an Escherichia coli/Enterococcus faecalis shuttle vector. The host range, transformation efficiency and stability of the Escherichia coli/Enterococcus faecalis shuttle vector were also studied. The results showed that plasmid pXW was 8617 bp in size and its GC content was 33.29%. It contained 8 ORFs and was assumed to be θ-type replicating plasmid. The copy number of plasmid pXW in E. faecalis EXW27 was up to 32.09±0.93, indicating that pXW was a high copy number plasmid. In this study, the minimal replicon of plasmid pXW was determined, and an Escherichia coli/Enterococcus faecalis shuttle vector was constructed based on this replicon. The shuttle vector had a wide host range and was successfully transformed into different types of lactic acid bacteria. The transformation efficiency was between 1.96×10²~8.96×10⁴ CFU/μg (plasmid DNA), and the plasmid loss rate was between 28.54% and 54.17%. In this study, we successfully constructed an Escherichia coli/Enterococcus faecalis shuttle vector with wide host range, high transformation efficiency and high stability, which provides a new tool for gene manipulation of lactic acid bacteria.
- Enterococcus faecalis,
- endogenous plasmid,
- θ-type replication,
- shuttle vector,
- Escherichia coli

FullText(HTML)

References (30)

References

[1]	DE FILIPPIS F, PASOLLI E, ERCOLINI D. The food-gut axis: Lactic acid bacteria and their link to food, the gut microbiome and human health[J]. FEMS Microbiology Reviews,2020,44(4):454−489.
[2]	WANG C, SHI C, ZHANG Y, et al. Microbiota in fermented feed and swine gut[J]. Applied Microbiology and Biotechnology,2018,102(7):2941−2948.
[3]	MOKOENA M P. Lactic acid bacteria and their bacteriocins: Classification, biosynthesis and applications against uropathogens: A mini-review[J]. Molecules,2017,22(8):1255.
[4]	KLEEREBEZEM M, KUIPERS O P, SMID E J. Lactic acid bacteria-a continuing journey in science and application[J]. FEMS Microbiology Reviews,2017,41(Supp_1):S1−S2.
[5]	KLAENHAMMER T R. Get cultured: Eat bacteria[J]. Annual Review of Food Science and Technology,2019,10:1−20.
[6]	PLAVEC T V, BERLEC A. Safety aspects of genetically modified lactic acid bacteria[J]. Microorganisms,2020,8(2):297.
[7]	PENG K, KOUBAA M, BALS O, et al. Recent insights in the impact of emerging technologies on lactic acid bacteria: A review[J]. Food Research International,2020:109544.
[8]	CHEN W, GU Z. Genomic analysis of lactic acid bacteria and their applications[M]. Singapore: Springer, 2018: 21-49.
[9]	PETERBAUER C, MAISCHBERGER T, HALTRICH D. Food-grade gene expression in lactic acid bacteria[J]. Biotechnology Journal,2011,6(9):1147−1161.
[10]	LANDETE J M. A review of food-grade vectors in lactic acid bacteria: From the laboratory to their application[J]. Critical Reviews in Biotechnology,2017,37(3):296−308.
[11]	DUONG T, MILLER M J, BARRANGOU R, et al. Construction of vectors for inducible and constitutive gene expression inLactobacillus[J]. Microbial Biotechnology,2011,4(3):357−367.
[12]	CUI Y, HU T, QU X, et al. Plasmids from food lactic acid bacteria: Diversity, similarity, and new developments[J]. International Journal of Molecular Sciences,2015,16(6):13172−13202.
[13]	SUZUKI K, SHINOHARA Y, KURNIAWAN Y N. Role of plasmids in beer spoilage lactic acid bacteria: A review[J]. Journal of the American Society of Brewing Chemists,2020,79(1):1−16.
[14]	CHO S W, YIM J, SEO S W. Engineering tools for the development of recombinant lactic acid bacteria[J]. Biotechnology Journal,2020,15(6):1900344.
[15]	MORRONI G, BRENCIANI A, LITTA-MULONDO A, et al. Characterization of a new transferable MDR plasmid carrying the pbp5 gene from a clade B commensal Enterococcus faecium[J]. Journal of Antimicrobial Chemotherapy,2019,74(4):843−850.
[16]	JENSEN L B, GARCIA-MIGURA L, VALENZUELA A J S, et al. A classification system for plasmids from Enterococci and other Gram-positive bacteria[J]. Journal of Microbiological Methods,2010,80(1):25−43.
[17]	FANG L S, LAI Q, ZHONG Z M, et al. Sequence analysis of an endogenous plasmid in Lactobacillus plantarum and construction of a shuttle expression vector using it[J]. Food Science,2020,41(4):118−124.
[18]	ZUO F, FENG X, SUN X, et al. Characterization of plasmid pML21 of Enterococcus faecalis ML21 from koumiss[J]. Current Microbiology,2013,66(2):103−105.
[19]	GREEN M R, SAMBROOK J. Molecular cloning: A laboratory manual[M]. New York: Cold Spring Harbor Laboratory Press, 2012.
[20]	CHEN Z, LIN J, MA C, et al. Characterization of pMC11, a plasmid with dual origins of replication isolated from Lactobacillus casei MCJ and construction of shuttle vectors with each replicon[J]. Applied Microbiology and Biotechnology,2014,98(13):5977−5989.
[21]	FRIESENEGGER A, FIEDLER S, DEVRIESE L A, et al. Genetic transformation of various species of Enterococcus by electroporation[J]. FEMS Microbiology Letters,1991,79(2−3):323−328.
[22]	WANG C, CUI Y, QU X. Optimization of electrotransformation (ETF) conditions in lactic acid bacteria (LAB)[J]. Journal of Microbiological Methods,2020:105944.
[23]	LANDETE J M, ARQUÉS J L, PEIROTÉN Á, et al. An improved method for the electrotransformation of lactic acid bacteria: A comparative survey[J]. Journal of Microbiological Methods,2014,105:130−133.
[24]	TERÁN L C, CUOZZO S A, ARISTIMUÑO FICOSECO M C, et al. Nucleotide sequence and analysis of pRC12 and pRC18, two theta-replicating plasmids harbored by Lactobacillus curvatus CRL 705[J]. PloS one,2020,15(4):e0230857.
[25]	KIM S W, JEONG E J, KANG H S, et al. Role of RepB in the replication of plasmid pJB01 isolated from Enterococcus faecium JC1[J]. Plasmid,2006,55(2):99−113.
[26]	WYCKOFF H A, BARNES M, GILLIES K O, et al. Characterization and sequence analysis of a stable cryptic plasmid from Enterococcus faecium 226 and development of a stable cloning vector[J]. Applied and Environmental Microbiology,1996,62(4):1481−1486.
[27]	WANG Y. Spatial distribution of high copy number plasmids in bacteria[J]. Plasmid,2017,91:2−8.
[28]	MARTÍNEZ-BUENO M, VALDIVIA E, GÁLVEZ A, et al. pS86, a new theta-replicating plasmid from Enterococcus faecalis[J]. Current Microbiology,2000,41(4):257−261.
[29]	LILLY J, CAMPS M. Mechanisms of theta plasmid replication[J]. Plasmids:Biology and Impact in Biotechnology and Discovery,2015,3(1):33−44.
[30]	SHERBA J J, HOGQUIST S, LIN H, et al. The effects of electroporation buffer composition on cell viability and electro-transfection efficiency[J]. Scientific Reports,2020,10(1):1−9.

[1]	HU Xiaoxia, DENG Lijuan, LIU Ruiting, SHI Rongmei. Evaluation of Medicinal Quality of Garlic Based on Principal Component Analysis[J]. Science and Technology of Food Industry, 2023, 44(12): 293-299. DOI: 10.13386/j.issn1002-0306.2022080237
[2]	HUANG Libiao, YUAN Yiyang, CHEN Lin, YANG Meiyan, PENG Zhiyuan, LU Xiaoting, GAO Xiangyang. Comprehensive Evaluation of Quality Characteristics of Different Mango Varieties Based on Principal Component Analysis and HS-SPME-GC-MS Technology[J]. Science and Technology of Food Industry, 2023, 44(3): 297-306. DOI: 10.13386/j.issn1002-0306.2022040221
[3]	Juanyuan HE, Xiang YU, Yanli FENG, Runfeng ZHANG, Lizhong CHEN, Hao HUANG. Muscle Quality Characterization and Principal Component Analysis of Deqing Shrimp[J]. Science and Technology of Food Industry, 2021, 42(8): 264-270. DOI: 10.13386/j.issn1002-0306.2020060342
[4]	GAO Yun, YU Zhi-fang. Quality Evaluation of Celery Based on Principal Component Analysis[J]. Science and Technology of Food Industry, 2020, 41(3): 308-314,320. DOI: 10.13386/j.issn1002-0306.2020.03.051
[5]	WEI Lu-lu, QIN Li-kang, WEN An-yan, ZHU Yi. Quality Evaluation of Different Varieties Millet Based on Principal Components Analysis[J]. Science and Technology of Food Industry, 2019, 40(9): 49-56. DOI: 10.13386/j.issn1002-0306.2019.09.010
[6]	LUO Hong-xia, WANG Li, JU Rong-hui, JIA Hong-liang, PAN Yan, LIN Shao-hua, ZHU Jian-chen. Quality Characteristics and Principal Component Analysis of Different Varieties of Millet Starch[J]. Science and Technology of Food Industry, 2018, 39(24): 11-17. DOI: 10.13386/j.issn1002-0306.2018.24.002
[7]	CAI Fang-yuan, WANG Pei, WANG Zhi-ting, LIU Jing, JIANG Mei. Evaluation of Rice Tofu Quality Based on Correlation Analysis and Principal Component Analysis[J]. Science and Technology of Food Industry, 2018, 39(17): 33-39,45. DOI: 10.13386/j.issn1002-0306.2018.17.006
[8]	YANG Ting, LEI Pan-deng, ZHOU Han-chen, DING Yong, CUI Peng, CHENG Man-huan, HUANG Jian-qin. Studies on aroma components in Taiping Houkui tea by HS-SPME-GC-MS coupled with principal component analysis[J]. Science and Technology of Food Industry, 2017, (10): 49-53. DOI: 10.13386/j.issn1002-0306.2017.10.001
[9]	LIU Sha-sha, ZHANG Bao-shan, SUN Xiao-yuan, LUO Teng. Principal components analysis of flavor compositions in Zizyphus jujube[J]. Science and Technology of Food Industry, 2015, (20): 72-76. DOI: 10.13386/j.issn1002-0306.2015.20.006
[10]	LUO Yan, RUAN Jun-xiang, SU Zhi-heng, LIN Cui, QIN Yang. Discrimination of liubao tea by FTIR and principal component analysis[J]. Science and Technology of Food Industry, 2014, (12): 55-57. DOI: 10.13386/j.issn1002-0306.2014.12.002

Supplements (0)

Cited By

Cited by

Periodical cited type(8)

1.	彭剑飞，施慧，李儒婷，郭苗苗，陈丽霞. 药食同源中药干预糖尿病及其并发症作用机制的研究进展. 河南中医. 2023(04): 625-630 .
2.	相欢，崔春. 沙棘籽粕蛋白肽的稳定性及分离纯化. 食品科学. 2023(18): 49-57 .
3.	王迪，李文霞，姚瑜，袁芳廷，袁木荣，彭强. 沙棘蛋白和多肽的提取及功能活性研究进展. 食品工业科技. 2022(03): 447-455 . 本站查看
4.	邢丽颖，李建颖，孙怡，韩东. 小浆果多肽研究进展. 粮食与油脂. 2022(02): 21-24 .
5.	赵轶轩，王丽娜，屈凝伊. 沙棘果研究进展. 中国民族民间医药. 2022(03): 56-62 .
6.	李文娟，初悦雷，潘雨欣，付王威，景智，胡勋矫，姚于飞，陈璿瑛. 白扁豆多糖通过HPA轴介导降血糖的作用机制. 食品工业科技. 2022(07): 361-367 . 本站查看
7.	刘均，李强，谭蓉. 基于糖代谢异常斑马鱼模型评价代用茶沙棘叶的降糖作用. 中国茶叶加工. 2022(01): 79-84 .
8.	杨明翰，盛萍. 新疆药食同源资源开发研究与前景展望. 中国实验方剂学杂志. 2021(13): 234-243 .