Lin28 B gene of duolang sheep and application of Lin28 B gene

A technology of Duolang sheep and genes, which is applied in the fields of application, genetic engineering, plant gene improvement, etc., can solve the problems that the regulation of puberty initiation in sheep is unknown, and there are no research reports on Lin28B gene cloning and function of Duoliang sheep. , to achieve huge potential economic value, change sexual maturity time, and improve reproductive performance

Inactive Publication Date: 2018-12-21
TARIM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The Lin28B gene was first cloned from humans. At present, the Lin28B genes of humans, mice, cattle, horses, cats, and pigs have been cloned, but so far, there has been no research on the cloning and function of the Lin28B gene of Duolang sheep. reported, but its regulatory effect on the onset of puberty in sheep is still unknown

Method used

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  • Lin28 B gene of duolang sheep and application of Lin28 B gene
  • Lin28 B gene of duolang sheep and application of Lin28 B gene
  • Lin28 B gene of duolang sheep and application of Lin28 B gene

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Amplification of the Lin28B gene fragment, template recovery, ligation transformation, and sequencing.

[0019] According to the provided sample tissue, the total RNA was extracted by TRIZOL method, and the cDNA reverse transcription kit Maxima of Thermo TM H Minus cDNA Synthesis Master Mix (M1662) reverse transcription, the specific steps are as follows.

[0020] 1. PCR amplification

[0021] 1) PCR reaction system

[0022] Component

Volume(μl)

2×SG PCR MasterMix

25

dna

1

Forward Piemr (10μM)

1

Reverse Primer (10μM)

1

ddH2O

22

Total

50

[0023] 2) PCR reaction program

[0024]

[0025] 3) 1% agrose gel electrophoresis, after confirming that there is specific amplification, then amplify two PCRs of 50 μl system, and recover the PCR products.

[0026] 2. Recovery of PCR products

[0027] 1) Add 4 times the volume (800μl) of Buffer CP to a 1.5ml centrifuge tube (containing a PCR reaction...

Embodiment 2

[0054] The present invention relates to the expression level of Lin28B gene of Duolang sheep in the hypothalamus / pituitary ovary of Duolang sheep.

[0055] 1. Total RNA extraction

[0056] 1) After grinding about 10-30mg of the sample with liquid nitrogen, transfer it to an RNase-free 1.5ml centrifuge tube, add 1ml of SGTRIzol lysate; shake vigorously, and let stand at room temperature for 2-3min;

[0057] 2) Add 200 μl chloroform, vortex vigorously for 30 seconds, and let stand at room temperature for 5 minutes;

[0058] 3) Centrifuge at 12000×g for 15 minutes at 4°C;

[0059] 4) Carefully transfer the supernatant to an RNase-free 1.5ml centrifuge tube, add an equal volume of isopropanol and mix well;

[0060] 5) Add 500 μl RNA Wash Buffer I to the column, centrifuge at 12000×g for 30s at 4°C. discard the filtrate;

[0061] 6) Centrifuge at 13000×g for 10 minutes at 4°C;

[0062] 7) discard the supernatant, add 1ml 70% ethanol to rinse the precipitate;

[0063] 8) Centr...

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PUM

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Abstract

The invention discloses a Lin28 B gene of duolang sheep and application of the Lin28 B gene. The Lin28 B gene is cloned from duolang sheep tissues and regulates the initiation of puberty; the Lin28 Bgene has two nucleotide sequences shown as SEQ ID NO:1 and SEQ ID NO:2 in the sequence table and can be applied to regulate the puberty initiation of the duolang sheep. The Lin28 B gene of the duolangsheep, disclosed by the invention, can artificially shorten the puberty age of sheep from heredity, and has very important significance and great potential economic value for the development of sheepfarming industry in China.

Description

technical field [0001] The patent of the invention belongs to the field of animal genetic engineering, and specifically relates to the Lin28B gene of Duolang sheep. Background technique [0002] Ewe puberty is the age when normal ewes show estrus for the first time and ovulate, and it is a sign of reproductive ability, and heredity is the main factor affecting ewe puberty. At present, the research on puberty initiation focuses on humans, mice and monkeys, and there are few studies on puberty initiation in sheep. In recent years, genome-wide association analysis (GWAS) and transgenic studies have found that the Lin28B gene plays an important role in regulating human puberty and the onset of puberty in mammals such as mice and goats, but its role in the onset of puberty in sheep is still unclear. None reported. The Lin28B gene was first cloned from humans. At present, the Lin28B genes of humans, mice, cattle, horses, cats, and pigs have been cloned, but so far, there has bee...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/47
CPCC07K14/47
Inventor 邢凤高庆华祁鑫李闯李庆瑾
Owner TARIM UNIV
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