Method for preparing human serum amyloid A1 and expression vector and genetic engineering bacteria thereof

A technology of genetically engineered bacteria and expression vectors, applied in the field of preparation of human serum amyloid A1 and its expression vectors and genetically engineered bacteria, can solve the problems of complicated purification methods, long time-consuming, low yield, etc., and achieve simplification of purification methods , avoid denaturation, low purification cost
CN101724641AInactive Publication Date: 2010-06-09DIASYS DIAGNOSTIC SYST SHANGHAI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
DIASYS DIAGNOSTIC SYST SHANGHAI
Publication Date
2010-06-09
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a method for preparing soluble genetic engineering human serum amyloid A1 (SAA1) and an expression vector and genetic engineering bacteria thereof. The expression vector provided by the invention is a plamid vector pET28a(+) containing a human SAA (hSAA1) gene sequence, and the hSAA1 encoding sequence is positioned between restriction enzyme cutting sites of NdeI and Xhol I on the pET28a(+) vector. The genetic engineering bacteria provided by the invention are colibacillus pET28a (+)-hSAA1 / Rosetta-gami, which can express soluble target protein. The preparation method of the invention uses a nickel cross-linking affinity-column one-step method, has simple operation, high yield and short consumed time, and is suitable for industrial large-scale production, and the purity of the obtained target protein is larger than 90%.
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Description

technical field

[0001] The present invention relates to the field of producing protein or polypeptide by DNA recombination technology, more specifically, the present invention relates to the method for preparing human serum amyloid A1 (Serum Amyloid A1, SAA1) through genetic engineering, and also relates to its expression vector and genetically engineered bacteria. Background technique

[0002] Serum Amyloid A (SAA) is an acute phase response apolipoprotein, and the SAA gene is located on chromosome 11, which is a group of pleomorphic proteins encoded by the same cluster of genes. Human serum amyloid A1 (SAA1) consists of 104 amino acids with a molecular weight of about 12KD-14KD. SAA is a polymorphic protein consisting of several related protein families (SAA1-SAA4). Mainly synthesized by liver cells. Among them, the genes of SAA1 and SAA2 have only 7 amino acid differences, and they both encode the acute phase SAA protein. SAA4 is little changed in the acute phase respo...

Claims

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