Transgenic yeast containing flounder growth hormone gene, preparation and application thereof

A growth hormone and transgenic technology, applied in the field of molecular biology technology and bioengineering, can solve problems such as disease and death, speed up the growth rate of flounder and flounder fish, and achieve the effects of promoting growth, improving feed conversion rate and low cost

Inactive Publication Date: 2008-10-08
OCEAN UNIV OF CHINA
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The invention provides a flounder growth hormone gene-transformed yeast and its preparation method and application, which can accelerate the growth rate of flounder and flounder fish, and solve problems caused by temperature, water quality and other reasons in the current long breeding process of flounder and flounder fish. The problem of sickness and death

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Transgenic yeast containing flounder growth hormone gene, preparation and application thereof
  • Transgenic yeast containing flounder growth hormone gene, preparation and application thereof
  • Transgenic yeast containing flounder growth hormone gene, preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1: Preparation of yeast with transgenic flounder growth hormone gene:

[0061] [1] Preparation of flounder growth hormone cDNA:

[0062] Extract total RNA from flounder pituitary gland, design primers ping1 and ping2 containing enzyme cutting sites BamH I and Not I, the ping1 primer sequence has 100% homology with the nucleotide sequence shown in No. 5 sequence of the sequence table, The ping2 primer sequence has 100% homology with the nucleotide sequence shown in the No. 6 sequence of the sequence table, and the flounder growth hormone cDNA is amplified by reverse transcription polymerase chain reaction (RT-PCR) method, and sequenced Make sure it matches the expected sequence.

[0063] [2] Construction of Saccharomyces cerevisiae transformation vector:

[0064] The flounder growth hormone mature peptide cDNA was digested with BamH I and Not I and then ligated into the Escherichia coli plasmid pUC-PGK to construct the transformation vector pIPGK-GH. The Es...

Embodiment 2

[0103] Embodiment 2: Identification of the yeast with transgenic flounder growth hormone gene:

[0104] Southern hybridization detection of flounder growth hormone gene recombination integrated into Saccharomyces cerevisiae genome DNA:

[0105] The genomic DNA of yeast transfected with flounder growth hormone gene was extracted, digested with EcoR I and BglI respectively, and the cDNA of flounder growth hormone was labeled with digoxin to make probes, and then Southern hybridization was performed. The result is as figure 2 Shown, wherein, M: λ / EcoRI+HindIII; 1: Genomic DNA of untransformed Saccharomyces cerevisiae (not digested); 2: Genomic DNA of untransformed Saccharomyces cerevisiae digested by EcoR I; Genomic DNA of untransformed Saccharomyces cerevisiae; 4: Genomic DNA of transformed Saccharomyces cerevisiae (undigested); 5: Genomic DNA of transformed Saccharomyces cerevisiae digested with EcoR I; 6: Genomic DNA of transformed Saccharomyces cerevisiae digested with Bgl ...

Embodiment 3

[0106] Example 3: Stability detection of flounder growth hormone gene in Saccharomyces cerevisiae:

[0107] Saccharomyces cerevisiae containing the flounder growth hormone gene was spread on the solid medium of Minimal Dextrose Plates without tryptophan, and after the colonies grew out, two clones were randomly selected and placed on the solid medium of Minimal Dextrose Plates without tryptophan Streak culture on the top; after the colonies grow out, randomly select 2 clones and culture them on the YPD solid medium... After 50 generations of continuous passage, then randomly select 10 colonies for PCR detection, all of which have specific stripes The results showed that the flounder growth hormone gene was stable in Saccharomyces cerevisiae and would not be lost with the passage of cells.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses recombinant yeast for the flounder growth hormone gene and the preparation method and the application thereof. The recombinant yeast is obtained through transferring the flounder growth hormone gene into saccharomyces cerevisiae, and the obtained saccharomyces cerevisiae expresses the flounder growth hormone. The fermentation technology of the saccharomyces cerevisiae is mature, the expression quantity is higher, the saccharomyces cerevisiae is secure and non-toxic, the obtained saccharomyces cerevisiae for expressing the flounder growth hormone is not required to be purified, and can be directly used as feedstuffs or feed additive for the flatfish cultivation, the growth speed of the flatfish can be quickened, and the cultivation cycle can be shortened.

Description

technical field [0001] The invention belongs to the field of molecular biology technology and bioengineering technology, in particular, it relates to a flounder growth hormone gene-transformed yeast and its preparation method and application. Background technique [0002] Since the commercial breeding of flounder in my country began in the 1980s, its proportion in aquaculture has been increasing because of its high market value and good economic benefits. By 2006, the aquaculture output of flounder in my country has exceeded 82,560 tons, and the output value has exceeded 6 billion yuan. Among them, flounder and turbot are currently the most important species in the aquaculture of flounder in my country. In recent years, Atlantic summer flounder, Desert flounder, Senegal sole, etc., the scale of artificial breeding of flounder is expanding day by day. However, during the long breeding process of flounder, due to the influence of temperature, water quality and other reasons, i...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N1/19C12N15/18C12N15/81C07K14/61A23K1/16A23K1/18C12R1/865A23K10/16A23K50/80
Inventor 张学成刘滨臧晓南刘顺梅
Owner OCEAN UNIV OF CHINA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap