Clone and expression of 7,8-desaturase gene
A dehydrogenase and gene technology, applied in the fields of genetic engineering and enzyme engineering, can solve the problems of low efficiency, difficult to control conditions, and many side reactions, and achieve the effects of reducing environmental pollution, shortening the process route, and improving the conversion efficiency.
Inactive Publication Date: 2009-08-19
GUANGXI UNIV
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Problems solved by technology
But so far, no matter what kind of chemical synthesis method is used to prepare raw material 7-dehydrocholesterol, there are disadvantages such as long reaction steps, low overall yield, many side reactions, difficult conditions to control, low efficiency, and high cost, thus affecting vitamin D. 3 production cost, sales price, marketing
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Embodiment 1
[0063] 1 Library sources and plasmids
[0064] Both the Drosophila cDNA library and pSE380 are preserved by our laboratory.
[0065] 2 main reagents
[0066] The pGM-T Simple Ligation Kit was purchased from Tiangen Biochemical Technology Co., Ltd., and the BioSpin Gel Recovery Kit and BioSpin PCR Product Purification Kit were purchased from Hangzhou Bioer Technology Co., Ltd. Both EcoR I and BamH I enzymes were purchased from Fermentas.
[0067] 3 methods
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Abstract
The invention provides clone and expression aiming at 7,8-dehydrogenase gene. The gene encoding 7,8-dehydrogenase is acquired from a constructed drosophila cDNA library, wherein the 7,8-dehydrogenase can transform cholesterol into 7-dehydrocholesterol, thereby providing a new method for synthesizing the 7-dehydrocholesterol by a biological process, acquiring a process route capable of shortening vitamin D3, reducing production cost, and reducing environmental pollution.
Description
technical field [0001] The invention belongs to the fields of genetic engineering and enzyme engineering, and specifically relates to the cloning and expression of 7,8-dehydrogenase genes. 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, maintain the balance of blood calcium and blood phosphorus, and can be used as pharmaceutical preparations, food and feed additives, etc., and can also be used clinically to treat rickets , senile osteoporosis, hypothyroidism, etc. current vitamin D 3 It is produced by photochemical method, that is, 7-dehydrocholesterol (7-DHC) is used as raw material, and it is converted into vitamin D after ultraviolet irradiation. 3 Precursor (PreD 3 ), obtained by reheating isomer...
Claims
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IPC IPC(8): C12N15/53C12N15/63C12N1/00C12P19/34C12N15/70C12R1/19
Inventor 黄时海李湘萍吴孔阳夏小斌陈桂光梁智群张云开李灿明汪晟康超何鑫平
Owner GUANGXI UNIV
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