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Stability analysis method for Taenia solium (TS) recombinant Lactococcus lactis expression system

A technology of Lactococcus lactis and Taenia solium, applied in the direction of recombinant DNA technology, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problems of recombination of Taenia solium that have not been seen yet

Inactive Publication Date: 2017-11-21
ZUNYI MEDICAL UNIVERSITY
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Problems solved by technology

[0004] As far as the above-mentioned background technical field is concerned, there have been no reports on the recombinant L.lactis of Taenia solium

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  • Stability analysis method for Taenia solium (TS) recombinant Lactococcus lactis expression system
  • Stability analysis method for Taenia solium (TS) recombinant Lactococcus lactis expression system
  • Stability analysis method for Taenia solium (TS) recombinant Lactococcus lactis expression system

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Embodiment Construction

[0032] The present invention will be further described below in conjunction with the embodiments and accompanying drawings, but the present invention is not limited to the following embodiments, and it can be foreseen that those skilled in the art may produce various changes in the implementation situation in combination with the prior art.

[0033] 1 Target gene analysis

[0034] According to the TSOL18 gene sequence of Taenia solium (accession number: AF017788.1), the target gene was analyzed and optimized.

[0035] 2 TSOL18 gene synthesis and primer design

[0036] Using the PAS (PCR-based Accurate Synthesis) method, Sac I and Hind III restriction sites and protective bases were designed at both ends of the primers, and TSOL18 and SP-TSOL18 were synthesized by Nanjing Zhongding Biological Laboratory target gene.

[0037] 3 Construction and identification of recombinant plasmids pMG36e-TSOL18 and pMG36e-SP-TSOL18

[0038] The synthesized target gene was ligated with the e...

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Abstract

The invention provides a stability analysis method for a Taenia solium (TS) recombinant Lactococcus lactis expression system. According to the stability analysis method, gene optimization is carried out by taking a TSOL18 gene sequence which is logged in the GenBank as a template and taking the Lactococcus lactis MG1363 as a host-based system to synthetize target genes TSOL18 and SP-TSOL18 so as to build expression vectors and expression systems. According to PCR identification and SDS-PAGE and Western blot analysis, intracellular target protein expression of TSOL18 is detected while extracellular target protein is not detected, but intracellular and extracellular target protein expressions of SP-TSOL18 are both detected; plasmids of pMG36e-TSOL18 and pMG36e-SP-TSOL18 are subjected to 20 times of passages continuously in L.lactis MG1363, and accordingly have good genetic stability. Thus, an effective reference base is provided for development and utilization of cysticercosis cellulosae vaccines.

Description

technical field [0001] The invention relates to the field of research on recombinant live vaccines of Taenia solium, in particular to a stability analysis method for a recombinant Lactococcus lactis expression system of the TSOL18 gene of Taenia solium. Background technique [0002] Cysticercosis cellulosae (cysticercosis cellulosae), commonly known as pig cysticercosis (bladder disease), is a serious hazard to animal husbandry production caused by the middle tapered larva cysticercus cellulosae of Taenia solium (Ts). Zoonosis, which is related to human health, is one of the key parasitic diseases planned and controlled by the Ministry of Health. The disease is distributed worldwide and is mainly prevalent in countries and regions with poor economic and sanitation conditions such as Central Africa, South Africa, Latin America, East Asia and South Asia. my country is mainly popular in 31 provinces, municipalities and autonomous regions including Heilongjiang, Jilin, Yunnan, ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/74C12N15/66C12N15/12
CPCC07K14/43554C12N15/66C12N15/746
Inventor 周必英孙俊超李想罗波江楠
Owner ZUNYI MEDICAL UNIVERSITY