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CR1 gene related to cell reconstitution, conduction and death

A technology of CR-1 and apoptosis, applied in genetic engineering, plant gene improvement, recombinant DNA technology, etc.

Inactive Publication Date: 2003-05-14
MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there has been no report on the correlation between a single gene and cell remodeling, conduction and apoptosis

Method used

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  • CR1 gene related to cell reconstitution, conduction and death
  • CR1 gene related to cell reconstitution, conduction and death
  • CR1 gene related to cell reconstitution, conduction and death

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0007] The cloning of embodiment 1CR-1 gene:

[0008] The present invention adopts a special cDNA cloning method [Life Sciences 63 (17): 1555-1562, 1998], that is, comprehensively adopts technologies such as cDNA selection, EST positioning, genomic DNA sequence analysis, PCR reaction and clone end extension, according to The combination of several elements involved searched NCBI's EST (expressed sequence tag) database, and the results showed that there were two EST clones (AX014823) (AX013073) containing 671 base pairs, and the sequence was consistent with any known protein The sequences were all different, and then two primers were synthesized,

[0009] Primer A: AGCGCGGTGAAGCGGGGGTGGGATCTG (SEQ ID No1)

[0010] Primer B: ATAACTTTATTTGCCTTTGGTGG (SEQ ID No2)

[0011] A human skeletal muscle cDNA library (purchased from Clontech) was used as a template, and primers A and B were used for PCR reaction. The PCR condition was 93°C for 2 minutes, followed by 30 cycles of 93°C fo...

Embodiment 2

[0017] Example 2 Detects the expression of CR-1 protein in various tumor cells by Western hybridization: use 5ml RPMI-1640 (Gibco company product) culture solution to cultivate cos7 African green monkey kidney epithelium respectively in a 25ml cell culture flask in a carbon dioxide incubator Cells, 7721 human liver cancer cells, 7402 human liver cancer cells, HT29 human rectal cancer cells, A549 human lung cancer cells, K562 human chronic leukemia cells, until the coverage rate reached about 90%, the cells were collected in 5ml pre-cooled 1×PBS (this Various reagents and conventional operation methods in the examples were prepared according to the instructions in the second edition of "Molecular Cloning Experiment Guide", and the cells were collected by centrifugation at 3000rpm for 5 minutes. Add 1:1 cell volume freshly prepared lysate and incubate on ice for 1 hour. Centrifuge at 10,000 rpm at 4°C for 15 minutes to collect the supernatant in a new tube, and quantify the prot...

Embodiment 3

[0018] Example 3 Using RNA to interfere with the expression of CR-1 gene in tumor cells, causing apoptosis of tumor cells:

[0019] RNA interference (RNAi) is a double-stranded RNA that induces sequence-specific post-transcriptional gene silencing, thereby inhibiting gene expression.

[0020] Synthetic oligodeoxynucleotides:

[0021] 5'GATCCGAGCCACAGCTAACAAGGCTTTTTTAAGCCTTGTTAGCTGTGGCTCT 3' (SEQ ID NO 5)

[0022] 5' CTAGAGAGCCACAGCTAACAAGGCTTAAAAAAGCCTTGTTAGCTGTGGCTCG 3' (SEQ ID NO 6)

[0023] A pair of primers were mixed and annealed, and cloned into the pBSKU6 vector (Liu Jing et al. High Technology Communication, Vol. 10, No. 8, pp. 6-9, 2000) and (Ren-Huan Xu BRAIN RESEARCH 899, 10-19).

[0024] Synthetic primers:

[0025] Primer C: 5'ATAACGCGTCCCGGGTGGTAAACCGTGCA3' (SEQ ID NO7)

[0026] Primer D: 5'TTAACGCGTGAGCTCAAGGTCGGGCAGGAA3' (SEQ ID NO8)

[0027] The U6 segment embedded with the RNAi fragment was amplified by PCR from the constructed pBSKU6 vector, and subclone...

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Abstract

The present invention relates to a human functional gene, especially CR-1 gene related to cell reconstitution, conduction and death. The gene is obtained through the comprehensive technological process of cDNA selection, EST location, genome DNA sequence analysis, PCR reaction and cloning end expanding, etc. Experiments including Northern and Western cross breeding detection of CR-1 gene expression in tumor cell, RNA interference immunohitochemistry detection of the expressino of CR-1 in myocardial tissue of myocardial disease patient, systemic research of the expression of CR-1 interacted protein and CR-1 gene in coliform bacteria and euaryotice cell, etc. show that the gene may be used as the target site in clinical diagnosis and treatment of myocardial disease and tumors.

Description

Technical field: [0001] The present invention relates to a human functional gene, especially a gene related to cell remodeling, conduction and apoptosis, and clarifies the regulation mechanism of the gene, especially its role in cardiomyocyte remodeling, tumor cell proliferation and apoptosis, It can provide effective means for the diagnosis and prevention of myocardial and tumor diseases. Background technique: [0002] Cell remodeling, conduction and apoptosis are related to many human diseases. For example, it is currently believed that cardiomyocyte remodeling and apoptosis play an important role in heart failure; cell information conduction is related to apoptosis and tumor occurrence. The study of genes or regulatory genes related to cell remodeling, conduction and apoptosis is of great significance for the diagnosis and treatment of human diseases, especially myocardial diseases and tumors, such as myocardial development genes MyoD and Myogenin regulate the proliferati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61P9/00A61P35/00C07H21/04C07K16/18C12N15/12C12N15/63C12Q1/68G01N33/574
Inventor 王以光李天伯金由辛胡洋李斌冯爽包慧中
Owner MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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