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Long-chain RNA Lnc-13814 for regulating and controlling follicle development of laying ducks and application thereof

A technology of rnalnc-13814 and follicle development, applied in the field of genetic engineering, to achieve the effect of improving the fecundity of laying ducks

Active Publication Date: 2019-10-01
INST OF ANIMAL SCI & VETERINARY HUBEI ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the research on tissue-specific LncRNAs in laying ducks, especially the follicular LncRNAs in laying ducks and its regulatory mechanism has not been reported.

Method used

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  • Long-chain RNA Lnc-13814 for regulating and controlling follicle development of laying ducks and application thereof
  • Long-chain RNA Lnc-13814 for regulating and controlling follicle development of laying ducks and application thereof
  • Long-chain RNA Lnc-13814 for regulating and controlling follicle development of laying ducks and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] 1. Laying duck long-chain RNA Lnc-13814 gene

[0037] The cDNA sequence corresponding to the laying duck long-chain RNA Lnc-13814 described in this example is shown in SEQ ID NO: 1. The laying duck long-chain RNA Lnc-13814 of the present invention is based on the full transcription of the laying duck follicle tissue completed by the applicant in the previous stage. Group sequencing results, a new LncRNA molecule obtained through a large number of bioinformatics analysis and screening of the sequencing results of two groups of follicular tissue samples at different developmental stages. The applicant named it the laying duck long-chain RNA Lnc-13814, and the length of the cDNA sequence corresponding to the laying duck long-chain RNA Lnc-13814 is 974bp.

[0038] 2. Preparation of the full-length sequence fragment of the long-chain RNA Lnc-13814 gene of laying ducks

[0039] 1. Sample collection

[0040] Collect 3 samples of egg-laying duck white follicles and 3 samples ...

Embodiment 2

[0071] Example 2 Fluorescent quantitative detection kit and method for laying duck long-chain RNA Lnc-13814 gene

[0072] In this example, a pair of fluorescent quantitative PCR detection primers was designed to identify and identify the objective presence of the laying duck long-chain RNALnc-13814 described in Example 1 in the laying duck follicular granulosa cells. The identification method of this embodiment includes the following steps:

[0073] 1. Sample collection

[0074] Collect 3 samples of egg-laying duck white follicles and 3 samples of yellow follicles during the peak laying period. After cleaning with PBS, puncture the follicles with a needle to squeeze out the follicle fluid, put them into an enzyme-free 1.5mL EP tube, mark them and put them into liquid nitrogen or Store in -80°C refrigerator.

[0075] 2. Extraction of total RNA from samples

[0076] Utilize traditional TRIzol, chloroform method to extract follicle tissue total RNA, specific steps are the same...

Embodiment 3

[0089] Example 3 Effect of long-chain RNA Lnc-13814 gene overexpression on apoptosis of laying duck follicular granulosa cells

[0090] 1. Construction of long-chain RNA Lnc-13814 gene overexpression vector in laying ducks

[0091] This embodiment provides an overexpression vector of the laying duck long-chain RNA Lnc-13814 gene, and the vector includes the cDNA sequence corresponding to the laying duck long-chain RNA Lnc-13814. Vector construction specifically includes the following steps:

[0092]1. Double digestion treatment of pcDNA3.1(+) vector

[0093] Schematic diagram of the structure of the pcDNA3.1(+) vector figure 1 As shown, the vector contains restriction endonucleases KpnI and XhoI cutting sites. The above vector was double-digested with two endonucleases KpnI and XhoI. The enzyme digestion reaction system was as follows, and the total volume of the system was 50 μL:

[0094] Table 4

[0095]

[0096] The reaction conditions are: enzyme digestion at 37°C ...

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Abstract

The invention provides a long-chain RNA Lnc-13814 for regulating and controlling follicle development of laying ducks and its application. A cDNA sequence corresponding to the long-chain RNA Lnc-13814related to follicular development of laying ducks is shown as SEQ ID NO:1. The invention also provides a fluorescent quantitative PCR detection primer pair of the long-chain RNA Lnc-13814 and detection kit. The detection primer pair is nucleotide sequences shown as SEQ ID NO:2 and SEQ ID NO:3. The invention also provides an over-expression vector of the long-chain RNA Lnc-13814 and a constructionmethod thereof. It is found for the first time that the long-chain RNA Lnc-13814 has the function of promoting the proliferation of follicular cells of laying ducks and has the important significancein study of the genetic nature of reproductive traits of laying ducks and improvement of the reproductive capacity of the laying ducks.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a long-chain RNALnc-13814 regulating egg follicle development of laying ducks and its application. Background technique [0002] Follicle development is closely related to the laying performance of laying ducks and directly affects the egg production of laying ducks. Studies have shown that poultry follicle development is an extremely complex biological process. At present, people have a certain understanding of the follicle development model of poultry. However, as an important factor that determines egg production, the specific regulatory mechanism of follicle development still needs to be further studied. Research. The growth of granulosa cells directly affects the development of follicles. Follicular granulosa cells can secrete gonadal hormones and growth factors to regulate the growth, differentiation and maturation of follicular theca cells and oocytes, and als...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12Q1/6888C12Q1/6851C12N15/11C12N15/85
CPCC12N15/113C12N15/85C12N2310/10C12N2800/106C12Q1/6851C12Q1/6888C12Q2600/124C12Q2600/158C12Q2600/178C12Q2531/113C12Q2545/114C12Q2563/107
Inventor 吴艳皮劲松肖红卫梁振华张昊潘爱銮蒲跃进申杰杜金平
Owner INST OF ANIMAL SCI & VETERINARY HUBEI ACADEMY OF AGRI SCI
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