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Liposome-encapsulated recombinant veterinary growth-promoting protein as well as preparation method and application of liposome-encapsulated recombinant veterinary growth-promoting protein

A growth-promoting and liposome technology, applied in the field of gene recombination technology and pharmaceuticals, can solve the problems of inability to take oral administration, loss of biological activity, etc., and achieve the effects of short half-life, wide application and low cost.

Active Publication Date: 2021-04-30
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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  • Liposome-encapsulated recombinant veterinary growth-promoting protein as well as preparation method and application of liposome-encapsulated recombinant veterinary growth-promoting protein
  • Liposome-encapsulated recombinant veterinary growth-promoting protein as well as preparation method and application of liposome-encapsulated recombinant veterinary growth-promoting protein
  • Liposome-encapsulated recombinant veterinary growth-promoting protein as well as preparation method and application of liposome-encapsulated recombinant veterinary growth-promoting protein

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 recombinant veterinary growth-promoting protein:

[0035] Refer to GENBANK accession number X53325.1, take the full-length amino acid sequence of porcine growth hormone, refer to GENBANK accession number M35609.1, take the full-length amino acid sequence of chicken growth hormone, and add the enzyme cleavage 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 restriction enzyme cleavage site EcoRI is added downstream, and the sequence is optimized. The target gene sequence shown in NO:1, wherein the flexible linker sequence is shown in SEQ ID NO:2.

[0036] At the same time, take the full-length amino acid sequence of the above-mentioned porcine growth hormone and the full-length amino a...

Embodiment 2

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

[0046] Five healthy male SD rats (purchased from Anhui Experimental Animal Center) were selected, weighing about 200 grams. After fasting for 12 hours, the liposome-encapsulated recombinant growth-promoting protein for veterinary use prepared in Example 1 was diluted to 1 mg / ml, and each rat was given 1 ml by gavage. Before the start of the test (0h) and 1, 3, 5, 7, 12, 18, and 24 hours after gavage, 200 μL of blood was collected from the tail of the mice, placed at 4 °C for 1 h, centrifuged at 7000 rpm for 15 minutes to separate serum, and immediately frozen at -70 °C , after the last blood collection, it was also frozen at -70°C for more than 2 hours to ensure that all sera were frozen and thawed once during the ELISA test. This recombinant protein is a fusion protein of porcine growth hormone and chicken growth hormone. Choose...

Embodiment 3

[0049] Example 3: Growth-promoting performance verification 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, and mix by mixing. The feeds are general-purpose feed for pigs and general-purpose feed for chickens produced by Anhui Xinkang Feed Factory. The feed added with L-VRGP 0.1 mg / g was recorded as feed A, the normal feed without L-VRGP was recorded as feed B, and the feed added with VRGP 0.1 mg / g was recorded as feed C.

[0051] Thirty white-feather chicks, 21-day-old and weighing 250±50g, half male and half male, were randomly divided into 3 groups, with 10 chickens in each group, as footmarks. Select 30 healthy ternary piglets, 60 days old and weighing 20±0.5kg, half male and half female, randomly divided into 3 groups, 10 pigs in each group, and ear tagging. Group A: 10 21-day-old white-feather chic...

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Abstract

The invention relates to the technical field of gene recombination, and particularly discloses lipidosome-encapsulated recombinant growth-promoting protein for animals as well as a preparation method and application of the lipidosome-encapsulated recombinant growth-promoting protein for animals, the nucleotide sequence of a coding gene of the recombinant growth-promoting protein for animals is shown as SEQ ID NO: 1, and the recombinant growth-promoting protein is fusion protein of pig growth hormone and chicken growth hormone and can simultaneously play a role in promoting growth of pigs and chickens; the molecular weight of the protein is large, and the problem of short half-life period can be well solved; the recombinant fusion protein is encapsulated in the liposome, so that decomposition of gastrointestinal tract protease can be avoided, and more importantly, the liposome has an effect of promoting absorption and can guide intestinal absorption of the protein. The veterinary growth-promoting hormone is produced in a recombination mode. Compared with an extraction method, the method is simple and low in cost and can be widely applied to feed additives.

Description

technical field [0001] The invention belongs to the technical field of gene recombination and the field of medicaments, in particular to a liposome-encapsulated recombinant veterinary growth-promoting protein (Liposome-coated veterinary recombinant growth-promoting protein, L-VRGP) and a preparation method and application thereof. Background technique [0002] Liposomes, also known as artificial membrane vesicles, are when amphiphilic molecules are dispersed in the water phase, the hydrophobic tails of the molecules tend to aggregate together, avoiding the water phase, while the hydrophilic heads are exposed in the water phase, forming a bilayer structure. The closed vesicles are non-toxic and non-immunogenic, and can be used for transgenic or drug preparation. [0003] Prokaryotic expression is a method category of protein recombination, which means that through gene cloning technology, the exogenous target gene is expressed in a specific prokaryotic organism or cell by con...

Claims

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Application Information

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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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