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Transformant for reducing weight and fat and construction method and application thereof

A transformant and lipid-lowering technology, applied in the field of biomedicine, can solve problems such as host drug resistance and host intestinal flora disorder, and achieve the effects of overcoming high production costs, convenient medication, and low cost

Active Publication Date: 2016-11-23
GUANGZHOU SINOGEN PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If taken for a long time, it may cause the host intestinal flora disorder, and also cause the host to develop drug resistance

Method used

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  • Transformant for reducing weight and fat and construction method and application thereof
  • Transformant for reducing weight and fat and construction method and application thereof
  • Transformant for reducing weight and fat and construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Preparation of Lactococcus lactis transformed with OXM gene.

[0041] (1) Construction of homologous recombination fragments:

[0042] According to the published thyA gene sequence in the whole genome of lactic acid bacteria L. lactis, the sequences of 1000 bp upstream of the start codon and 1000 bp downstream of the stop codon were respectively found.

[0043] According to the human OXM amino acid and nucleic acid sequence, referring to the codon usage frequency of Lactococcus lactis, the OXM nucleic acid sequence was optimized, and the obtained sequence is shown in SEQ ID NO:1.

[0044] Artificial chemically synthesized gene fragment OXM-M: Lactococcus lactis thyA gene start codon upstream 1000bp + human OXM gene sequence + Lactobacillus thyA stop codon downstream 1000bp sequence.

[0045] (2), preparation and screening of OXM transgenic Lactococcus lactis:

[0046] The human oxyntomodulin fragment OXM-M was electrotransformed into the strain L.lactis NZ9...

Embodiment 2

[0065] Example 2: Pharmacodynamic experiment of OXM-transformed lactic acid bacteria for weight loss and lipid-lowering.

[0066] (1) Several newly weaned B6 mice were prepared and fed with high fat until the obese mice were successfully established. The obesity standard is: (body weight of the experimental group-average body weight of normal mice) / average body weight of normal mice>20%. The obese mice were divided into two groups, 10 in each group, and continued to be fed with a high-fat diet. One group is the control group, fed with Lactococcus lactis L. lactis NZ9000. The other group is the experimental group, feeding Lactococcus lactis L. lactis NZ9000-OXM. At the same time, a normal control group was set up, that is, normal diet B6 mice were not fed Lactococcus lactis.

[0067] (2) Each mouse was given 5*10 by gavage 9 CFU Lactococcus lactis, orally administered once every 2 days. The body weight, food intake and other vital signs of the mice were tracked and observe...

Embodiment 3

[0072] Example 3: Pharmacodynamic experiment of OXM-transformed lactic acid bacteria on weight loss and lipid-lowering in gene-deficient diabetic mice.

[0073] 1. Prepare 20 db / db mice at the age of 8 weeks, and divide the obese mice into 2 groups, 10 in each group. One group is the control group, fed with Lactococcus lactis L. lactis NZ9000. The other group is the experimental group, fed with Lactococcus lactis L.lactisNZ9000-OXM. At the same time, a normal control group was set up, that is, normal diet B6 mice were not fed Lactococcus lactis.

[0074] 2. Each mouse was given 5*10 by gavage 9 CFU Lactococcus lactis, orally administered every day. Weight monitoring data such as Figure 11 As shown, the body weight of mice fed with OXM group can be maintained unchanged compared with their own body weight before administration. However, the body weight of the mice in the NZ9000 group continued to increase by about 8% in 5 weeks. Through this experiment, it can be found th...

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Abstract

The invention discloses a transformant for reducing weight and fat, the transformant is obtained by recombination and replacement of human oxyntomodulin gene and thymidylate synthetase gene in lactococcus lactis genome, and the recombination and replacement comprises artificially chemically synthesizing gene sequences, to be more specific, allowing both ends of the human oxyntomodulin gene to comprise homology ends of both ends of the thymidylate synthetase gene in the lactococcus lactis genome, and electrically transferring gene segments into lactococcus lactis for homologous recombination. The transformant for reducing weight and fat is capable of constitutive expression of oxyntomodulin in genome, does not contain a resistance gene, can safely and effectively reduce weight, fat and blood sugar, and can be used for preparing medicines for the treatment of obesity, diabetes and lipid metabolism disorder.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a transformant for weight loss and lipid reduction, its construction method and application. Background technique [0002] Obesity is a health issue of great concern in the world today. In developed countries in Europe and America, more than half of the people are overweight, and 20-30% of them are obese patients. Obesity not only affects movement and quality of life, but also can cause a series of diseases such as cardiovascular disease, type 2 diabetes and kidney disease, which are extremely harmful to human health. Survey statistics show that for every unit increase in body mass index (BMI), the risk of developing hypertension increases by 10%. [0003] Although there are many causes of obesity, factors such as genetics, endocrine disorders, diet structure and lifestyle can all induce obesity. However, the vast majority of obese patients are simple obesity caused by poor diet and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/21C12N15/90C12N15/16A23L1/29A61K48/00A61K38/26A61P3/04A61P3/06A61P3/10C12R1/01
CPCA61K8/64A61K35/744A61K38/26A61Q19/06A61P1/16A61P3/04A61P3/06A61P3/10C07K14/605C12N9/1007C12N15/902A61K35/74A61K38/22A61K38/27C07K14/575C12N15/907
Inventor 林艳向荣赵子建李芳红
Owner GUANGZHOU SINOGEN PHARMA CO LTD
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