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Vector, cell and method for improving developmental rate of yak cloned embryos

A technology for cloning embryos and developmental rates, applied in chemical instruments and methods, genetically modified cells, botany equipment and methods, etc., to achieve the effect of improving in vitro developmental rates

Active Publication Date: 2022-03-18
NORTHWEST A & F UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most domestic animals have been successfully cloned through somatic cell nuclear transfer technology, but so far the yak, an important domestic animal species in the Qinghai-Tibet region, has not been successfully cloned

Method used

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  • Vector, cell and method for improving developmental rate of yak cloned embryos
  • Vector, cell and method for improving developmental rate of yak cloned embryos
  • Vector, cell and method for improving developmental rate of yak cloned embryos

Examples

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Embodiment 1

[0046] 1. The primary culture of yak fetal fibroblasts

[0047] From the Yanzi Slaughterhouse of Xining City, Qinghai Province, the yak embryo, the yak embryo, the yak fetus, the eluted yak fetus, completely extracted with the fetal membrane, with penicillin (200 IU / mL), streptomycin (200 IU / mL) After 500 ml of physiological saline, rapidly washed 3 times, transferred to the thermosette (adding dodge-containing physiological saline), rapidly taking back to the laboratory in half an hour.

[0048] Tissue block culture method culture yak fetus fibroblasts:

[0049] In the sterile ultimate platform, the yak fetus is placed in a sterilized beaker, and the saline preheated saline preheated in advance is cleaned 3-5 times. Sterilization Ophthalmology, lightly peeled off the skin tissue of the fetus, shifts in 60 mm culture In the dish, PBS liquid cleans 3 times, sterilized eye-cut cut skin tissue to 1 mm 3 Size, DMEM high-sugar culture liquid rehabilitation tissue, 1200r / min centr...

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Abstract

The invention discloses a vector, a cell and a method for improving the developmental rate of a yak cloned embryo, and belongs to the technical field of yak cloned embryos. The vector is an eukaryotic expression vector of shRNA (short hairpin ribonucleic acid), the shRNA is targeted to a CBX3 gene, and a target sequence of the shRNA is shown as SEQ ID NO. 1: GGTCTTGATCCAGAACGAATA. Transfecting yak somatic cells through the vector to obtain positive monoclone; the positive monoclone is used as a donor to carry out nuclear transfer and construct the cloned embryo, so that abnormal reprogramming of the yak cloned embryo can be corrected, the in-vitro development rate of the yak cloned embryo is remarkably improved, and a material basis and a theoretical basis are provided for clarification of a reprogramming mechanism of the yak cloned embryo.

Description

Technical field [0001] The present invention belongs to the field of yakclon embryos, and in particular to a carrier, cell, and method for increasing the development rate of yakclon embryos. Background technique [0002] BOS GRUNNIENS lives in the Himalayas and Qinghai-Tibet Plateau in the Himalayan and Qinghai-Tibet Plateau, Bovinae. Because it can accommodate the harsh ecological environment such as high-precipitation, hypoxia and strong radiation, there is a "boat of the plateau", which is one of the local important livestocks, which provides local farmers and herdsmen and other necessities such as meat, milk and fur. Can be used as a local important transportation tool. However, the yak reproduction and low growth performance, and the high-quality breeding livestock is greatly limited to the development of animal husbandry. [0003] Through body cell nuclear transfer technology (i.e. cloning technology), oocytes can reprogram the body cells to melt-embryonic cells and ultimat...

Claims

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Application Information

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IPC IPC(8): C12N15/85C12N15/113C12N5/10C12N15/877
CPCC12N15/85C12N15/113C12N5/0656C07K14/47C12N15/8771C12N2310/14C12N2510/00C12N2800/107
Inventor 苏建民余彤张成图吴英孟茹陈永忠
Owner NORTHWEST A & F UNIV
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