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A liposome-encapsulated recombinant veterinary growth-promoting protein and its preparation method and application

A growth-promoting and liposome technology, which is applied in the field of gene recombination technology and the field of pharmaceuticals, can solve the problems of being unable to take it orally and losing biological activity, and achieve the effects of short half-life, wide application, and low cost

Active Publication Date: 2021-11-30
安徽中起生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, relevant institutions have developed recombinant porcine growth hormone, chicken growth hormone, etc., but most of them cannot be taken orally. Some companies have also developed porcine growth hormone fusion protein containing transducin, but this oral protein has the effect of protease in the gastrointestinal tract. It will be decomposed soon and lose its original biological activity

Method used

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  • A liposome-encapsulated recombinant veterinary growth-promoting protein and its preparation method and application
  • A liposome-encapsulated recombinant veterinary growth-promoting protein and its preparation method and application
  • A liposome-encapsulated recombinant veterinary growth-promoting protein and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Preparation of liposome-encapsulated recombinant veterinary growth-promoting protein

[0034] (1) Acquisition of the gene sequence of the recombinant veterinary growth-promoting protein:

[0035] Refer to GENBANK accession number X53325.1 to obtain the full-length amino acid sequence of porcine growth hormone, and refer to GENBANK accession number M35609.1 to obtain the full-length amino acid sequence of chicken growth hormone, and add the enzyme cutting site BamHI, porcine growth hormone sequence and chicken growth hormone upstream A flexible linker is used between the sequences to protect the spatial structure of the target fragment, and the enzyme cutting site EcoRI is added downstream, and the sequence is optimized, and the sequence-optimized target gene sequence is sent to Huada Gene for synthesis, and the sequencing is qualified, such as SEQ ID The target gene sequence shown in NO:1, wherein the flexible linker sequence is shown in SEQ ID NO:2.

[0036...

Embodiment 2

[0045] Example 2: Detection of serum concentration of liposome-encapsulated recombinant veterinary growth-promoting protein after oral administration

[0046] Select 5 healthy male SD rats (purchased from Anhui Experimental Animal Center), weighing about 200 grams. After fasting without water for 12 hours, the liposome-encapsulated recombinant veterinary growth-promoting protein PBS prepared in Example 1 was diluted to 1 mg / ml, and each rat was given 1 ml by intragastric administration. Take 200 μL of blood from the tail of the rat before the test (0h) and 1, 3, 5, 7, 12, 18, and 24 hours after gavage, place at 4°C for 1 hour, centrifuge at 7000rpm for 15 minutes to separate the serum, and immediately freeze it at -70°C , After the last blood collection, it was also frozen and stored at -70°C for more than 2 hours to ensure that all serum was frozen and thawed once during ELISA testing. The recombinant protein is a fusion protein of porcine growth hormone and chicken growth h...

Embodiment 3

[0049] Example 3: Verification of growth-promoting performance of liposome-encapsulated recombinant veterinary growth-promoting protein

[0050] Take the liposome-encapsulated recombinant veterinary growth-promoting protein prepared in Example 1, add 10 g of L-VRGP to 100 kg of feed for mixing and mixing. The feeds were general-purpose feeds for pigs and general-purpose chickens produced by Anhui Xinkang Feed Factory. The feed added with 0.1 mg / g of L-VRGP was recorded as feed A, the common feed without L-VRGP was recorded as feed B, and the feed added with 0.1 mg / g of VRGP was recorded as feed C.

[0051] Select 30 white-feathered chicks aged 21 days and weighing 250±50g, half male and half male, randomly divided into 3 groups, 10 chickens in each group, and make foot marks. Select 30 healthy three-yuan piglets aged 60 days and weighing 20±0.5kg, half male and half female, randomly divided into 3 groups, 10 pigs in each group, and ear tagged. Group A: 10 21-day-old white-fe...

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Abstract

The present invention relates to the technical field of genetic recombination, and specifically discloses a liposome-encapsulated recombinant veterinary growth-promoting protein and its preparation method and application. The nucleotide sequence of the gene encoding the recombinant veterinary growth-promoting protein is shown as SEQ ID NO Shown in: 1, it is the fusion protein of porcine somatotropin and chicken somatotropin, which can act on the growth promotion of pigs and chickens simultaneously; the molecular weight of this protein is relatively large, and can better solve the problem of short half-life; adopt liposome Encapsulating the recombinant fusion protein in it can avoid the decomposition of protease in the gastrointestinal tract, and more importantly, the liposome has the function of promoting absorption and can guide the intestinal absorption of the protein. The invention uses recombination to produce animal growth-stimulating hormone, which is simpler and lower in cost than the extraction method, and can be widely used in feed additives.

Description

technical field [0001] The invention belongs to the technical field of gene recombination and the field of pharmaceuticals, and in particular relates to a liposome-encapsulated recombinant veterinary growth-promoting protein (Liposome-coated veterinary recombinant growth-promoting protein, L-VRGP) and its preparation method and application. Background technique [0002] Liposomes, also known as artificial membrane bubbles, are when amphiphilic molecules are dispersed in the water phase, the hydrophobic tails of the molecules tend to gather together and avoid the water phase, while the hydrophilic heads are exposed to the water phase to form a bilayer structure. The closed vesicles are non-toxic and non-immunogenic, and can be used for genetic modification or preparation of drugs. [0003] Prokaryotic expression is a method category of protein recombination. It refers to the method of constructing an expression vector and introducing an expression strain of an exogenous targe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70A61K38/27A61K9/127A61K47/24A61K47/28A61P3/00A23K20/147A23K50/30A23K50/75
CPCA61K9/127A61K38/00A61K47/24A61K47/28A23K20/147A23K50/30A23K50/75A61P3/00C07K14/61C07K2319/00C12N15/70
Inventor 卞传忠
Owner 安徽中起生物科技有限公司
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