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Novel gene Xsynuclein for xenopus laevis GRK family

A Xenopus laevis family technology, applied in genetic engineering, plant gene improvement, transferase, etc., can solve problems such as extraction of cDNA clone reports

Inactive Publication Date: 2008-05-28
SHANDONG UNIV
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Problems solved by technology

[0007] Although the synuclein series has great research and development value, there is no report on the extraction of the complete cDNA clone of Xsynuclein, and its spatiotemporal expression during development is also little known. The extraction method of this new gene, its encoded polypeptide , RT-PCR primer design, RT-PCR methods and whole-body in situ hybridization techniques are all new fields

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  • Novel gene Xsynuclein for xenopus laevis GRK family

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Embodiment Construction

[0030] The technical content of the embodiment of the present invention is specifically described below in conjunction with accompanying drawing:

[0031] Cloning, sequencing and expression of a novel gene Xsynuclein of the GRK family in Xenopus laevis

[0032] 1. Gene cloning

[0033] We constructed a phage cDNA library using ectoderm cells in the animal cap of scattered stage 8 Xenopus embryos treated with activin (Nieuwkoop PD, Faber J. Normal table of Xenopus laevis. North Holland: Amsterdam 1975; Cao Y, Zhao H , Hollemann T, ChenY, Grunz H. Tissue-specific expression of an ODC paralogue, XODC2, in Xenopus laevis[J]. Mech Dev, 2001, 102: 243-246), and then convert the phage cDNA library into plasmid cDNA by excision Library (Stratagene, USA). The library was screened using large-scale whole-mount in situ hybridization. We sequenced the newly screened clone, and performed sequence analysis in the online gene bank of the National Center for Biotechnology Information (NCBI...

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Abstract

The invention discloses the total length cloning of the cDNA of the new gene Xsynuclein of the GRK family of African laevis. The total length of the cDNA is 1040bp, which includes a complete and opening reading frame and can code the protein containing 129 amino acids. The invention also discloses two TR-PCR primers of the gene, TR-PCR method and technique of whole-mount in situ hybridization. The new gene Xsynuclein of the GRK family of African laevis has high level expression in the central nervous system in the period of embryo and adult. The new gene Xsynuclein can take part in the process of forming the forebrain of the embryo and has close relations to neurodegenerative diseases, breast cancer, ovarian cancer and the like. The new gene Xsynuclein of the GRK family of African laevis and the coding protein thereof will hopefully make breakthrough in curing neurodegenerative diseases, breast cancer, ovarian cancer and the like.

Description

technical field [0001] The present invention belongs to the field of molecular biology, and specifically relates to a Xenopus gene, and more specifically relates to a full-length cDNA clone of the new gene Xsynuclein of the Xenopus GRK family and its spatiotemporal expression during Xenopus embryonic development. The extraction method of a new gene, its nucleotide sequence and coded polypeptide, two RT-PCR primers, RT-PCR method and overall in situ hybridization technology. Background technique [0002] Neurogenesis is one of the earliest and most important stages in the process of animal morphogenesis. Studying the expression regulation of some important genes in the process of neurogenesis is very important for revealing the molecular mechanism of morphogenesis. G protein-coupled receptor kinases (GRK) mediate their biological responses by producing second messengers such as cAMP, and carry out their desensitization downstream by phosphorylating active receptors Regulati...

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Application Information

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IPC IPC(8): C12N15/54C12N15/10C12N9/12C07H21/00C12Q1/68A61K31/7088A61K38/45A61P35/00A61P25/28
Inventor 刘瑶屈庆春
Owner SHANDONG UNIV
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