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Nucleotide sequence of cDNA code area of goat Scp3 gene

A sequence and encoding technology, applied in the field of cDNA, can solve the problems of no report on Scp3 research, no report of goat Scp3 gene cDNA nucleotide sequence amino acid sequence, no report of goat Scp3 protein antibody, etc.

Inactive Publication Date: 2009-11-25
INNER MONGOLIA UNIVERSITY
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Problems solved by technology

[0005] So far, studies on the regulatory role of Scp3 in mammalian germ cell meiosis 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 Scp3 in goat cells. There is also no report on the cDNA nucleotide sequence of the goat Scp3 gene and its corresponding amino acid sequence, so there is no report on the antibody corresponding to the goat Scp3 protein

Method used

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  • Nucleotide sequence of cDNA code area of goat Scp3 gene
  • Nucleotide sequence of cDNA code area of goat Scp3 gene
  • Nucleotide sequence of cDNA code area of goat Scp3 gene

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Embodiment 1

[0026] Embodiment 1: Cloning of the cDNA coding region of goat Scp3 gene In order to clone the cDNA of the Scp3 coding region from goat testis tissue cells, according to the nucleotide sequence (NM_153694, NM_013041, NM_011517) disclosed in Figure 1 (A, B, C), Firstly, PCR degenerate primers allowing amplification of 985 bp cDNA were prepared, ie upstream primer 5'-GAAAAATGGTGCCCTCTGGAAG-3' and downstream primer 5'-GGAGGTGAACAAAAGAATGGT-3'.

[0027] In order to clone the cDNA of Scp3 by RT-PCR method, goat total RNA was isolated from goat testis tissue, and the total RNA of goat testis tissue was extracted by using Watson's Handbook for RNA Isolation & Purification version 2.07 according to the operating procedure of the instruction manual. For RNA, the concentration of RNA was determined by ultraviolet light and stored in a -80°C refrigerator for later use. Then, use TaKaRa's M-MLV reverse transcriptase and perform a reverse transcription reaction according to the operating p...

Embodiment 2

[0032] Embodiment 2: the nucleotide sequence of the cDNA of goat Scp3 gene

[0033] figure 2 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 are shown. exist figure 2 , the sequence shown is the nucleotide sequence of Scp3, which includes the 708 bp nucleotide sequence of the cDNA coding region, encoding 235 amino acids forming the mature protein with an inferred molecular weight of 27.7 KDa.

[0034]Figure 6 (A, B, C) shows the comparison of the nucleotide sequence of goat Scp3 gene cDNA with the nucleotide sequence of human (NM_153694), rat (NM_013041) and mouse (NM_011517) Scp3 cDNA. It shows that: 1) the nucleotide sequence of goat Scp3 gene cDNA has 89% homology with human Scp3 gene cDNA nucleotide sequence; it has 84% ​​homology with rat Scp3 gene cDNA nucleotide sequence; It has 89% homology with mouse Scp3 nucleotide sequence. The amino acid sequence deduced from this goat ...

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Abstract

The invention relates to a nucleotide sequence of a cDNA code area of a goat Scp3 gene and a corresponding coded amino acid sequence, in particular to the cDNA which is used for coding a Scp3 protein and separated from goat spermary cells and an amino acid sequence corresponding to the cDNA. In the invention, a conserved region is found through comparing the known nucleotide sequences of Scp3 genes of human, rat and mouse; a pair of degenerate primers for amplifying the cDNA code area of the goat Scp3 gene with a RT-PCR method is designed without breaking amino acid coding, and the pair of the degenerate primers is amplified to form a specific fragment through the RT-PCR method; and sequencing is carried out to obtain the nucleotide sequence of the cDNA code area of the goat Scp3 gene and the amino acid sequence corresponding to the nucleotide sequence. The obtained nucleotide sequence of 708 bp can be used for detecting the tissue expression specificity of the Scp3 gene through the RT-PCR method or a hybridization method, and can be used for preparing an Scp3 antibody after a protein is obtained by recombined expression.

Description

technical field [0001] The present invention relates to the cDNA encoding synaptonemal complex protein 3 (Scp3) isolated from goat testicular cells, more specifically to the entire nucleotide sequence of the cDNA coding region encoding synaptonemal complex protein 3 and its corresponding amino acid sequence . Background technique [0002] Meiosis is a special form of cell division. In meiosis, chromosomes and their DNA are replicated once, and cells divide twice in succession, resulting in haploid gamete cells. Meiosis is divided into meiosis I and meiosis II. Meiosis II is similar to normal mitosis, whereas meiosis I is a rather specialized process in which sister chromatids do not separate, but homologous chromosomes move to the poles of the cell separately. The completion of this process depends on many delicate subcellular structures and is accompanied by a series of complex events, including the pairing of homologous chromosomes, the assembly of synaptonemal complex ...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/47
Inventor 吴应积侯越罗奋华
Owner INNER MONGOLIA UNIVERSITY
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