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Goat S6K1 gene cDNA encoding zone nucleotide sequence

A nucleotide sequence, S6K1 technology, applied in the field of cDNA, can solve the problems affecting the survival rate of newborns, no S6K1 research report, no goat S6K1 gene cDNA nucleotide sequence, etc.

Inactive Publication Date: 2008-08-20
INNER MONGOLIA UNIVERSITY
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

It has been proved by experiments that the knockout of S6K1 (S6K1 - / - ) mice, the survival rate was significantly reduced, indicating that S6K1 affects the survival rate of newborns
[0004] So far, studies on the regulatory role of S6K1 in mammalian cell growth and proliferation have been carried out in human, mouse, bovine and other mammalian cells and many achievements have been made, but there is no research report on S6K1 in goat cells. There is no report on the cDNA nucleotide sequence of goat S6K1 gene and its corresponding amino acid sequence

Method used

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  • Goat S6K1 gene cDNA encoding zone nucleotide sequence
  • Goat S6K1 gene cDNA encoding zone nucleotide sequence
  • Goat S6K1 gene cDNA encoding zone nucleotide sequence

Examples

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Embodiment 1

[0021] Embodiment 1: the cloning of the cDNA coding region 1563bp fragment of goat S6K1 gene

[0022] In order to clone the cDNA containing the 1563bp coding region from the S6K1 gene of goat muscle tissue cells, according to the nucleotide sequence (AY396564) disclosed in Figure 1, first prepare the PCR primers that allow the amplification of the 1563bp cDNA, that is, the upstream primer 5'ATGAGGCGACGAAGGAGGC 3' and downstream primer 5'CAGGTGCTCTGGTCGTTTG 3'. Then add a BamHI restriction site and a protective base GC to the 5' end of the upstream primer, and add a stop codon TGA and an EcoRI restriction site and a protective base GC to the 5' end of the downstream primer, that is, an upstream primer P15'GC GGATCC ATGAGGCGACGAAGGAGGC 3', downstream primer P25'GC GAATTC TCA CAGGTGCTCTGGTCGTTTG 3'.

[0023] In order to clone the cDNA of S6K1 gene by RT-PCR method, total RNA was isolated from goat muscle tissue, and the total RNA of goat muscle tissue was extracted by using TaKa...

Embodiment 2

[0028] Embodiment 2: the nucleotide sequence of the cDNA of goat S6K1 gene

[0029] Fig. 2 shows the nucleotide sequence (SEQ ID NO: 1) of the cDNA clone obtained in Example 1, and the amino acid sequence (SEQ ID NO: 2) deduced therefrom. In FIG. 2, the sequence shown is the nucleotide sequence of the S6K1 gene, which includes the 1563 bp nucleotide sequence of the cDNA coding region, encoding 521 amino acids forming a mature protein with an inferred molecular weight of 58.6 KDa.

[0030] Fig. 6 shows the comparison of the nucleotide sequence of goat S6K1 gene cDNA with the nucleotide sequence of bovine (AY396564) S6K1 gene cDNA. It shows that: 1) the nucleotide sequence of goat S6K1 gene cDNA has 99% homology with the nucleotide sequence of bovine S6K1 gene cDNA. The amino acid sequence deduced from this goat nucleotide sequence has 2 amino acid differences compared with the amino acid sequence of the bovine S6K1 protein fragment at the same position (AAR01025), with a homol...

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Abstract

The invention relates to a cDNA nucleotide sequence which encodes S6K1 protein and is separated from a goat muscle cell and the corresponding amino acid sequence. The invention finds a conservation region by comparing the cDNA nucleotide sequences of the known human, rat, rabbit and cattle S6K1 genes and then designs a pair of primers for a cDNA coding area fragment of an RT-PCR amplified goat S6K1 gene according to the cDNA sequence of a cattle S6K1 gene (GenBank landing number is AY396564), the primers are used for amplifying the specific fragment by the PT-PCR method, the nucleotide sequence of 1563bp of the cDNA coding area of the goat S6K1 gene and the corresponding amino acid sequence are obtained after sequencing. The obtained 1563bp nucleotide sequence can be further used for full-length cDNA cloning of the goat S6K1 gene and detecting the tissue expression specificity of the S6K1 gene by the RT-PCR or the hybridization method, which can also be used for preparing an antibody for detecting the S6K1 after recombinant expression.

Description

technical field [0001] The present invention relates to the cDNA encoding S6K1 protein isolated from goat muscle cells, more specifically relates to the 1563bp nucleotide sequence of the cDNA coding region encoding the S6K1 protein and its corresponding amino acid sequence. Background technique [0002] S6K1 is a Ser / Thr kinase belonging to the AGC kinase family. The protein contains five main domains, which are NT (N-terminal domain), catalytic domain, the linker region, autoinhibitory domain and CT (C -terminal domain). In mammals, it is known that the S6K1 gene cDNA coding region of cattle consists of 1584 nucleotides, encoding 527 amino acid residues, and the S6K1 gene coding region of rabbits, humans and rats consists of 1578 nucleotides, encoding 525 amino acid residues. [0003] S6K1 is a positive regulator of cell growth and cell cycle progression. In the mTOR signaling pathway, it receives stimulation from the upstream mTOR (themammalian target of rapamycin), and ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/47
Inventor 旭日干吴应积王志钢
Owner INNER MONGOLIA UNIVERSITY
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