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Modified human serum albumin gene suitable for expression in silk gland of silkworm and its expression system and application

A technology of human serum albumin and expression system, which is applied in the expression system of human serum albumin gene and the field of expression of recombinant human serum albumin in the silk gland of silkworm, can solve the problems of high price and disease transmission, and achieve huge market development prospects and control The effect on the risk of disease transmission

Active Publication Date: 2018-05-11
SOUTHWEST UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the commercial production of human serum albumin is mainly realized through blood separation and purification, which is not only very expensive, but also has the risk of disease transmission, such as hepatitis and HIV

Method used

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  • Modified human serum albumin gene suitable for expression in silk gland of silkworm and its expression system and application
  • Modified human serum albumin gene suitable for expression in silk gland of silkworm and its expression system and application
  • Modified human serum albumin gene suitable for expression in silk gland of silkworm and its expression system and application

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Experimental program
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Effect test

Embodiment 1

[0033] 1. Gene synthesis

[0034] Download the amino acid sequence of the mature peptide of human serum albumin (Homo sapiens albumin, HSA, GI: 215982788) from NCBI, optimize the coding sequence according to the codon usage preference of silkworm, and put it at the C-terminal of the amino acid sequence of human serum albumin (HSA) Fusion of 6 histidine amino acids to form HSA-His6. For the convenience of operation, BamHI and NotI restriction sites are respectively connected to the two ends of the nucleotide sequence of HSA-His6, that is, the sequence shown in SEQ ID NO.1, The amino acid sequence encoded by the expression cassette is shown in SEQ ID NO.2.

[0035] 2. Construction of transgene expression vector

[0036] Artificially synthesized a double-strand containing the human serum albumin gene coding sequence (SEQ ID NO.1) designed through codon optimization, digested with BamHI and NotI to replace the psl1180[hr3Pser1spRedSer1PA] vector (see the publication number for th...

Embodiment 2

[0040] 1. Detection of recombinant human serum albumin (rHSA) content in total cocoon shell protein

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Abstract

The invention discloses a reconstructed human serum albumin gene suitable for cultivated silk gland expression and an expression system and application thereof. The reconstructed human serum albumin gene is as shown as the 7-1767th in SEQ ID NO.1, and the coded amino acid is as shown in SEQ ID NO.2. The reconstructed human serum albumin gene, a secretion type sericin 1 gene promoter and a secretion type sericin 1 gene terminator form an expression frame, an enhancer Hr3 is used for enhancing expression, and simultaneously a piggyBac transposon and a fluorescent screening marker gene construct an expression system. The expression system can highly effectively express recombined human serum albumin in cultivated silk gland, has biological activity, can be used for large scale production of recombined human serum albumin, and has good market prospect.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a modified human serum albumin gene, an expression system containing the modified human serum protein gene and a method for expressing recombinant human serum albumin by silk gland of silkworm silkworm. Background technique [0002] Since the beginning of the 21st century, with the increasing demand for functional proteins for various purposes such as medical, medicinal, edible, beauty, and health care, relying on extraction and production of proteins from natural sources has been unable to meet the rapidly growing market demand. . Establish and improve various high-efficiency prokaryotic and eukaryotic expression systems, and use E. coli, yeast, insect cells, mammalian cells, insects and mammals as host bioreactors to achieve low-cost, large-scale production with biological activity An effective and sustainable method for recombining foreign proteins, and has beco...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/14C07K14/765C07K1/36C07K1/34C07K1/22C12N15/85C12N15/89A01K67/04
CPCA01K67/0339A01K2207/15A01K2217/072A01K2227/706A01K2267/01C07K14/765C12N15/8509C12N15/89C12N2800/105C12N2800/22C12N2800/90C12N2830/008
Inventor 王峰夏庆友王元成王日远赵萍
Owner SOUTHWEST UNIV
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