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Recombinant plasmids for producing hydroxyl salidroside and genetically engineered bacteria and application thereof

A technology for the production of hydroxy red and recombinant plasmids, which is applied in genetic engineering, plant gene improvement, recombinant DNA technology, etc., can solve the unrecorded production of hydroxysalidroside, etc., and achieve broadening of innovative methods, low price and high product yield Improved effect

Inactive Publication Date: 2021-03-16
CHONGQING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The synthesis process needs to regulate glucose and other substrates, and the bacteria will be affected by metabolites, and there is no record of how to generate hydroxysalidroside in the future

Method used

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  • Recombinant plasmids for producing hydroxyl salidroside and genetically engineered bacteria and application thereof
  • Recombinant plasmids for producing hydroxyl salidroside and genetically engineered bacteria and application thereof
  • Recombinant plasmids for producing hydroxyl salidroside and genetically engineered bacteria and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0051] Example Synthesis of Hydroxysalidroside

[0052] The synthetic route of hydroxysalidroside is as follows: figure 1 As shown, it is mainly divided into the following seven steps:

[0053] 1. Preparation of culture medium

[0054] LB Medium: Contains 5g of yeast powder, 10g of tryptone and 10g of NaCl per liter of water. The solid medium is 1.5% agar powder added to the liquid medium. In order to increase the selectivity of the medium, the concentration of ampicillin (Amp) and kanamycin (Kan) was 100 μg / mL.

[0055] Yeast YPD medium: 1% yeast extract, 2% peptone, 2% glucose.

[0056] Yeast solid screening medium: medium solid powder SD (purchased from coolaber science), and 2% agar powder was added.

[0057] 2. Preparation of recombinant plasmids

[0058] First design primers:

[0059] Lys0X-F:5'-catg ccatgg atgacaatgttgactttgctgaat-3';

[0060] Lys0X-R:5'-cg ggtacc ttagatcttcacctggtactccttg-3';

[0061] gcd-F:5'-cgg ggtacc atggcaattaacaatacaggctcgc-3';

...

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Abstract

The invention relates to the field of bioengineering, and particularly discloses recombinant plasmids for producing hydroxyl salidroside and genetically engineered bacteria and application of the recombinant plasmids and the genetically engineered bacteria. According to the invention, tyrosine hydroxylase BvTH, AAS enzyme gene and glycosidase UGTs gene are respectively cloned to a plasmid vector pTrc99a to obtain recombinant plasmids PTAX4511 and RrTG1068, and then the recombinant plasmids are transferred into engineering bacteria NVSL4869; through overexpression of tyrosine hydroxylase BvTH,AAS enzyme gene and glycosidase UGTs gene, saccharomyces cerevisiae can obtain the capacity of producing target enzyme, so that the purpose of producing hydroxyl salidroside is achieved. The strain and plasmid disclosed by the invention can be used for producing a downstream derivative hydroxyl salidroside of an important drug molecule salidroside by taking tyrosine as a substrate under the condition of not influencing the survival of saccharomyces cerevisiae, so that the application of salidroside in pharmacology and medicine is broadened.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a recombinant plasmid for producing hydroxysalidroside, a genetically engineered bacterium and its application. Background technique [0002] Rhodiola has been used in the medical history of many countries. Modern research has found that Rhodiola can promote the ability of red blood cells to carry oxygen, has a good effect on altitude sickness, and has a good regulatory effect on the immune system. Rhodiola has multiple regulatory mechanisms such as lowering blood sugar, anti-oxidative stress, and inhibiting vascular smooth muscle proliferation. It can effectively treat diabetic vascular complications and is used for the treatment of various systemic diseases in the human body. The function of stress, as a traditional Chinese medicine adaptogen, has a certain regulatory and protective effect on human organs, so Rhodiola rosea is becoming more and more popular in the treatment of var...

Claims

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Application Information

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IPC IPC(8): C12N15/81C12N15/53C12N15/54C12N1/19C12P19/44C12R1/865
CPCC12N15/81C12N9/0071C12N9/1051C12P19/44C12Y114/16002C12Y204/01017
Inventor 王丹罗若诗杨锡志徐彦芹秦钊周丹高鑫张进许
Owner CHONGQING UNIV
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