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Polypeptide-human protein containing linking protein family characteristic sequential fragment-9.68 and polynucleotide for encoding it

A polynucleotide and protein family technology, applied in peptide/protein components, anti-animal/human immunoglobulin, DNA/RNA fragments, etc., can solve related protein expression and function abnormalities, metabolic or developmental disorders, etc. question

Inactive Publication Date: 2003-04-02
FUDAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The mutation or abnormal expression of the above two consensus sequence fragments will lead to abnormal expression and function of related proteins in the body, and then cause various related metabolic or developmental disorders

Method used

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  • Polypeptide-human protein containing linking protein family characteristic sequential fragment-9.68 and polynucleotide for encoding it
  • Polypeptide-human protein containing linking protein family characteristic sequential fragment-9.68 and polynucleotide for encoding it
  • Polypeptide-human protein containing linking protein family characteristic sequential fragment-9.68 and polynucleotide for encoding it

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Experimental program
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Embodiment 1

[0144] Total RNA was extracted from human fetal brain by one-step method of guanidine isothiocyanate / phenol / chloroform. Poly(A) mRNA was isolated from total RNA using Quik mRNA Isolation Kit (product of Qiegene). 2ug poly(A) mRNA was reverse transcribed to form cDNA. Smart cDNA cloning kit (purchased from Clontech) was used to insert the cDNA fragment into the multiple cloning site of the pBSK(+) vector (product of Clontech Company), transform DH5α, and the bacteria formed a cDNA library. The sequences of the 5' and 3' ends of all clones were determined using Dyeterminate cycle reaction sequencing kit (product of Perkin-Elmer) and ABI 377 automatic sequencer (Perkin-Elmer). Comparing the determined cDNA sequence with the existing public DNA sequence database (Genebank), it was found that the cDNA sequence of one of the clones, 1722c11, was a new DNA. The insert cDNA fragment contained in this clone was determined bidirectionally by synthesizing a series of primers. The resu...

Embodiment 2

[0144] Total RNA was extracted from human fetal brain by one-step method of guanidine isothiocyanate / phenol / chloroform. Poly(A) mRNA was isolated from total RNA using Quik mRNA Isolation Kit (product of Qiegene). 2ug poly(A) mRNA was reverse transcribed to form cDNA. Smart cDNA cloning kit (purchased from Clontech) was used to insert the cDNA fragment into the multiple cloning site of the pBSK(+) vector (product of Clontech Company), transform DH5α, and the bacteria formed a cDNA library. The sequences of the 5' and 3' ends of all clones were determined using Dyeterminate cycle reaction sequencing kit (product of Perkin-Elmer) and ABI 377 automatic sequencer (Perkin-Elmer). Comparing the determined cDNA sequence with the existing public DNA sequence database (Genebank), it was found that the cDNA sequence of one of the clones, 1722c11, was a new DNA. The insert cDNA fragment contained in this clone was determined bidirectionally by synthesizing a series of primers. The resu...

Embodiment 3

[0146] The human protein-9.68 containing the characteristic sequence fragment of the connexin family of the present invention has homology with the MOTIF domain, the homology result is shown in Figure 1, the homology rate is 20%, the score is 10.69; the threshold is 10.85. Example 3: Cloning the gene encoding human protein-9.68 containing the characteristic sequence fragment of the connexin family by RT-PCR

[0147] The total RNA of fetal brain cells was used as a template, and oligo-dT was used as a primer to carry out reverse transcription reaction to synthesize cDNA. After purification with a Qiagene kit, PCR amplification was performed with the following primers:

[0148] Primer1: 5'-CATCCTGAGAACTGAAATTGATCGC-3'(3)

[0149]Primer2: 5'-ATAAAATTTTTGAATTTATGTTCAA-3'(4)

[0150] Primer1 is the forward sequence starting from the 1bp at the 5' end of 1;

[0151] Primer2 is the 3' reverse sequence in 1.

[0152] The conditions of the amplification reaction: 50mmol / L KCl, 10mmo...

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Abstract

A polypeptide-human protein-9.68 containing vinculin family characteristic sequence fragment, the polynucleotide for coding it, the process for preparing said polypeptide by DNA recombination, the application of said polypeptide in treating diseases such as digestive ulcer, diabetes, etc, the antagonist of said polypeptide and its medical action, and the application of said polynucleotide are disclosed.

Description

technical field [0001] The invention belongs to the field of biotechnology. Specifically, the invention describes a new polypeptide-human protein-9.68 containing the characteristic sequence fragment of the junction protein family, and the polynucleotide sequence encoding the polypeptide. The present invention also relates to the preparation method and application of the polynucleotide and polypeptide. Background technique [0002] Connexins are eukaryotic proteins involved in regulating the interconnection of actin filaments with the plasma membrane in vivo. Junction proteins usually associate with the plasma membrane through the adhesion sites of the plasma membrane. In addition to actin, connexin is also combined with other structural proteins in organisms such as talin and α-actinin. [0003] Connexin is a large protein of about 116Kd containing an acidic N-terminal domain of about 90Kd and a C-terminal domain of about 25Kd of a proline-ric...

Claims

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Application Information

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IPC IPC(8): C07H21/04C07K14/435C07K14/47C07K16/18C12N15/10C12N15/11C12N15/113C12N15/12C12N15/63C12N15/64
Inventor 毛裕民谢毅
Owner FUDAN UNIV
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