Prawn Y-organ 7,8-dehydrogenase gene and application thereof

A dehydrogenase and gene technology, applied in the field of genetic engineering, can solve problems such as long process flow, easy pollution of the environment, and low yield, and achieve the effects of shortening the process route, reducing environmental pollution, and reducing production costs

Inactive Publication Date: 2015-04-29
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] So far, no matter what kind of chemical synthesis method is used to prepare the raw material 7-dehydrocholesterol, there are disadvantages such as long process flow, many st

Method used

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  • Prawn Y-organ 7,8-dehydrogenase gene and application thereof
  • Prawn Y-organ 7,8-dehydrogenase gene and application thereof
  • Prawn Y-organ 7,8-dehydrogenase gene and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment

[0033] 1. Library sources and plasmids

[0034] The Y-organ cDNA library and pPICZaA of Newpenaeus ensis (Metapenaeus ensis) are preserved by our laboratory.

[0035] 2. Main reagents

[0036] The expression host strain P. pastoris X-33 (wild type) and the antibiotic Zeocin were purchased from Invitrogen. Both Pmll enzyme and XbaI enzyme were purchased from TaKaRa Bioengineering Company.

[0037] 3. Method

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PUM

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Abstract

The invention belongs to the field of gene engineering, and particularly relates to a prawn Y-organ 7,8-dehydrogenase gene and application thereof. The invention provides a gene for coding 7,8-dehydrogenase, which is obtained from a constructed Metapenaeus ensis Y-organ cDNA (complementary deoxyribonucleic acid) library; and the nucleotide sequence of the gene is disclosed as SEQ ID NO.1. The application method comprises the following steps: cloning the 7,8-dehydrogenase coding gene, and connecting with an expression plasmid to construct a recombinant expression plasmid; after linearizing and purifying the recombinant expression plasmid, converting a receptor bacterium, and screening to obtain a gene engineering bacterium; and carrying out induction expression, purification and identification to obtain the 7,8-dehydrogenase of which the amino acid sequence is disclosed as SEQ ID NO.2. The 7,8-dehydrogenase can be used for converting cholesterol into 7-dehydrocholesterol by a one-step enzyme process, thereby enhancing the conversion efficiency, shortening the process route for producing vitamin D3, lowering the production cost and reducing the environmental pollution.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, in particular to a shrimp Y-organ 7,8-dehydrogenase gene and application thereof. Background technique [0002] Vitamin D 3 It is an essential fat-soluble vitamin for the growth, development, reproduction, maintenance of life and health of humans and animals. Its main function is to regulate calcium and phosphorus metabolism, promote intestinal calcium and phosphorus absorption and bone calcification, and maintain the balance of blood calcium and blood phosphorus. In addition, vitamin D 3 It is also a new type of immunomodulator, which has the functions of inducing cell differentiation, inhibiting cell growth, antagonizing collagen formation, regulating the expression of proto-oncogenes, and inhibiting tumor growth. Today, the aging of the world's population has become one of the social problems that attract worldwide attention. Both at home and abroad attach great importance to th...

Claims

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Application Information

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IPC IPC(8): C12N15/53C12N9/04C12N15/81C12P33/02C12R1/84
Inventor 黄时海苏辉兰王国盼李丽张光旭鄢凯舟盘慧群黄广上梁智群陈桂光刘诗宇李连威付跃
Owner GUANGXI UNIV
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