Non-resistance screening double antigen anchor expression vector plq2a and its preparation method

An expression vector and non-resistance technology, which is applied in the field of Lactobacillus plantarum non-resistance screening double antigen-anchored expression vector pLQ2a and its preparation field, can solve the problem of positive recombination in screening of exogenous plasmid electrotransformin host bacteria and Lactobacillus plantarum ligation products Difficulties in fertilization, cell wall thickness of Lactobacillus plantarum, etc.
CN108410901BActive Publication Date: 2020-06-30JILIN AGRICULTURAL UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
JILIN AGRICULTURAL UNIV
Publication Date
2020-06-30

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Abstract

The invention discloses a non-resistant plantarum lactobacillus anchor expression vector pLQ2a, the base sequence of which is shown in SEQ ID NO.1; it is a biosafety exogenous protein expression system, with asd deletion strain E. coli 6212 / Δasd and alr missing L. plantarum NC8 / Δalr is the host bacterium, which will be derived from Lactococcus lactis ( Lactococcus lactis subsp.cremoris strain JM4) The P23 promoter on the genome acts as the promoter of alr gene expression ， Using pYA4545, pYA4545‑Mcherry, pLP‑1261 Inv, pSIP409PgsA', pSIP409PgsA'‑IL‑10, pSIP409PgsA'‑EGFP as base vectors to construct nutrient complementation screening markers in Escherichia coli with PgsA' and LP‑1261 as dual anchor models ‑Lactobacillus has no anti-anchoring expression vector pLQ2a, and EGFP and Mcherry are used as reporter genes for screening and anchoring expression verification; it solves the problem of low conversion efficiency of Lactobacillus plantarum and it is difficult to screen positive recombinants, and constructs the expression vector pLQ2 both in E. coli 6212 / Δalr replicate expression, but also in L. plantarum Expressed in NC8 / Δalr.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a double-antigen-anchored expression vector pLQ2a for non-resistance screening of Lactobacillus plantarum and a preparation method thereof. Background technique

[0002] Lactobacillus plantarum ( Lactobacillus plantarum , L. plantarum ) as an intestinal host strain can tolerate the environment of low pH value and high salt in the stomach, the optimum growth temperature is 30-37 ℃, and can tolerate low temperature of about 10 ℃, but it appears in the environment above 45 ℃ Growth inhibition, commonly found in fermented foods made from cream, meat, vegetables, etc. and can be isolated from most plants, hence the name. Studies have shown that Lactobacillus plantarum complies with the GRAS (Generally Recognized as Safe) regulations of the US Food and Drug Administration (FDA) and the Qualified Presumption of Safety standards of the European Food Safety Authority (EFSA), an...

Claims

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