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Expression and purification method of schistosoma japonicum SjP40 protein and application

A technology for expression and purification of schistosomiasis, applied in chemical instruments and methods, peptide/protein components, medical preparations containing active ingredients, etc., to achieve the effect of easy operation and simple method

Active Publication Date: 2015-04-29
苏州美仑生物科技有限公司
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  • Abstract
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Problems solved by technology

However, as a mixed protein, SEA has not been reported on the effect of its component proteins, especially the P40 protein, on hepatic stellate cells

Method used

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  • Expression and purification method of schistosoma japonicum SjP40 protein and application
  • Expression and purification method of schistosoma japonicum SjP40 protein and application
  • Expression and purification method of schistosoma japonicum SjP40 protein and application

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Embodiment Construction

[0028] (1) Materials and methods

[0029] 1. Materials

[0030] The eggs of Schistosoma japonicum were purchased from Jiangsu Institute of Schistosomiasis Control, and the LX-2 cell line was purchased from the cell bank of Xiangya Central Laboratory of Central South University. Escherichia coli (E.coli.) DH5α, BL21, prokaryotic expression vector pET-28a (+) plasmid (Novagen company product), preserved by our laboratory; human recombinant TGF-β1 was purchased from sigma company.

[0031] Trizol reagent was purchased from Invitrogen Company of the United States; reverse transcription reagent was purchased from Fermentas Company of the United States; primers were synthesized by Shanghai Sangon Bioengineering Co., Ltd.; SYBR Premix Ex TaqTM Kit was purchased from Takara Company of Japan; mouse anti-human α-SMA, type I collagen and more The cloned primary antibody was purchased from Santa Cruz Company of the United States, the anti-GAPDH antibody was purchased from Hangzhou Xianzh...

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Abstract

The invention discloses an expression and purification method of schistosoma japonicum SjP40 protein and an application. The method comprises the steps of construction and identification of recombinant expression plasmid, primer design, adding of a restriction enzyme cutting site and a protective base to the two ends of the primer, PCR construction of pET-28a-SjP40 by taking pMD19-T-SjP40 as a template, expression and purification of recombinant plasmid and recombinant protein and the like. The invention also provides an application of the schistosoma japonicum P40 protein in preparing a preparation inhibiting the protein-level expression of alpha alpha-SMA and I-type collagen in the LX-2 cell and an application in preparing a preparation inhibiting the activation of TGF-beta1 induced LX-2 cells. The method disclosed by the invention is simple and easy to operate; and the invention provides a new application of the schistosoma japonicum P40 protein.

Description

technical field [0001] The invention relates to a method for expressing and purifying Schistosoma japonicum SjP40 protein. Background technique [0002] In the prior art, the expression and purification method of Schistosoma japonicum egg antigen SjP40 is relatively complicated, and the application of Schistosoma japonicum SjP40 protein (P40 protein of mainland China strain) has not been effectively developed, which has been a long-term research topic in this field. [0003] Schistosomiasis liver fibrosis is caused by schistosomiasis infection, mainly manifested as inflammatory granuloma formation centered on eggs and liver fibrosis. The formation of egg granuloma in the host liver and the occurrence and development of liver fibrosis will eventually cause the host to develop portal hypertension syndrome, complicated by upper gastrointestinal bleeding and hepatic coma, and eventually lead to the death of the host. Like liver fibrosis caused by other factors, the key link of ...

Claims

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Application Information

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IPC IPC(8): C07K14/44C12N15/70A61K38/17A61P33/12
CPCA61K38/00A61K38/17C07K14/44C12N15/70Y02A50/30
Inventor 段义农朱丹丹陈金铃孙晓雷冯金荣沈培何雪张令博
Owner 苏州美仑生物科技有限公司
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