Development and applications of new human gene FAMLF in human gene recombination, malignant tumor gene detection and specific monoclonal antibody

A malignant tumor and gene technology, applied in the field of medicine, can solve problems such as inhibition, malignant tumor, and continuous activation of cell proliferation signals

Inactive Publication Date: 2010-09-22
FUJIAN MEDICAL UNIV UNION HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The expression of the new gene FAMLF in patients with acute myeloid leukemia was significantly higher than that in normal people, and the difference was statistically significant (P<0.01). Bioinformatics analysis predicted that the new gene FAMLF might be located in the cell membrane and participate in cell information Phosphorylation of certain proteins in the delivery pathway; high expression of FAMLF may lead to continuous activation of cell proliferation signals or inhibition of cell apoptosis signals, resulting in the occurrence of malignant tumors

Method used

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  • Development and applications of new human gene FAMLF in human gene recombination, malignant tumor gene detection and specific monoclonal antibody
  • Development and applications of new human gene FAMLF in human gene recombination, malignant tumor gene detection and specific monoclonal antibody
  • Development and applications of new human gene FAMLF in human gene recombination, malignant tumor gene detection and specific monoclonal antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0179] Example 1: Cloning the full-length sequence of the FAMLF gene by sMART-RACE technology

[0180] 1) EST zywB-87 (GenBank registration number: CV973101) sequence proofreading

[0181] 1.1) Aligning EST zywB-87 with the genome

[0182] networking http: / / genome.ucsc.edu / cgi-bin / hgBlat? command=start Comparing EST zywB-87 with the human genome sequence, the following results are obtained:

[0183] Alignment of YourSeq and chr1: 197136247-197173157

[0184] Click on links in the frame to the left to navigate through the alignment. Matching bases in cDNA and genomic sequences are colored blue and capitalized. Light blue bases mark the boundaries of gaps in either sequence(often splicesites).

[0185] cDNA YourSeq

[0186] acgcggggAC AGGAGCAAGG GATGTCTGAG CACAAGTGGC TGAGTTCCGA 50

[0187] GTGACTTTAT GAAGCACTTT CTACCTTCCT CTCCGGCATG AAAACAGGGA 100

[0188] TTCTGCACCT GCATCATGGA CAGTCTGGCA AAAGCCTCTG CTCTGCCTCC 150

[0189] GGGGACAAGA AACTAGAGCA AATAACCGcT TTGAAATTAG ATC...

Embodiment 2

[0255] Example 2: Bioinformatics analysis and functional prediction of FAMLF

[0256] 1) Full-length cDNA analysis results

[0257] Apply the NCBI / ORF Finder program ( http: / / www.ncbi.nlm.nih.gov / gorf / gorf.html ) to analyze its ORF coding frame, the results are shown in Figure 7 . The code number of the sequence table is SEQ ID NO: 1, and the sequence of the cDNA of FAMLF is as follows:

[0258] 1 AGAACTGCAG ATAGTACAGC TTCCACAGGA GCAAGGGATG TCTGAGCACA AGTGGCTGAG 60

[0259] 61 TTCCGAGTGA CTTTATGAAG CACTTTCTAC CTTCCTCTCC GGCATGAAAA CAGGGATTCT 120

[0260] 121 GCACCTGCAT CATGGACAGT CTGGCAAAAG CCTCTGCTCT GCCTCCGGGG ACAAGAAACT 180

[0261] 181 AGAGCAAATA ACCGTTTTGA AATTAGATCC TGGCAAAATT ACCAACAATC AATGATGGAA 240

[0262] 241 ACTGAAGGGA GATTGATTTA AATTCTTCAAT GACAGTTGTA CAGAAGTTGA AGAAACACGT 300

[0263] 301 CATCTTTCCA AAGAAGAGTG AGTAGCTTGG TTGGTGGTAC CCTAAAATTA GGGCATGATT 360

[0264] 361 AAGGTACGTC ATTTCTTTTA GTTCAATCGA AGTGCTTTGC TGTTTCCATT TCAGAGAATT 420

[0265] 421 T...

Embodiment 3

[0329] Embodiment 3: Cloning of the full-length cDNA of human gene FAMLF

[0330] According to the sequence of the human gene FAMLF, a pair of primers F82 and R1974 were designed to amplify the full-length cDNA of the FAMLF gene including the signal peptide sequence. Amplification conditions: denaturation at 95°C for 15 minutes; The program cycled 35 times at 72°C for 30s (2min in total length); the final extension was 7min at 72°C. The PCR product was verified by direct sequencing and cloned into the vector pMD18-T. PCR and sequencing confirmed its correctness. The gel recovery kit of Shanghai Sangon Company was used for DNA fragment recovery, the plasmid extraction kit of Shanghai Sangong Company was used for plasmid extraction, and the heat shock method was used for Escherichia coli transformation. See "Molecular Cloning" for specific steps. DNA sequence determination showed that the DNA sequence of the PCR product was the same as 82-1974bp shown in Sequence Listing 1. ...

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Abstract

The invention discloses development and applications of a new human gene FAMLF in human gene recombination, malignant tumor gene detection and a specific monoclonal antibody, wherein, the new human gene FAMLF comprises cDNA of the new gene related to familial acute myelogenous leukemia and a polynucleotide sequence for completely opening a reading frame; the gene comprises a cDNA full-length sequence, wherein, the sequence table code thereof is SEQ ID No:1; and the sequence for completely opening the reading frame, wherein, the sequence table code thereof is SEQ ID No:2. The gene sets up the foundation for a diagnosis antibody, a protein chip, a gene chip and a gene medicine of the specific gene of leukemia in the future, thus playing an important role in developing and researching the new anti-leukemia medicine, improving diagnosis accuracy and treatment effect of leukemia and improving recovery rate of leukemia in Fujian province, and having potentially significant social and economic benefits. The invention lays the foundation for gene diagnosis and gene treatment of leukemia and other malignant tumors.

Description

Technical field: [0001] The invention relates to the field of medicine, in particular to the full-length DNA sequence of a novel human gene (FAMLF), the construction of a T clone of the full-length cDNA of the gene, the design of PCR primers for the sequence-specific amplification region of the gene, and the development of The specific monoclonal antibody against the protein expressed by the gene has been produced, and the subdivided fields are as follows: [0002] (1) The field of molecular biology. [0003] (2) Antibody preparation research field. [0004] (3) The field of clinical blood disease research. Background technique: [0005] With the completion of the Human Genome Project, human genomics research has achieved a comprehensive transformation from the structural genome to the post-genome, and the sequences of most of the 24 chromosomes in the human genome (except for a few intervals) have been clarified. It is estimated that there are approximately 20,000-25,000...

Claims

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Application Information

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IPC IPC(8): C12N15/12C12N15/11C12N15/09C07K14/47C07K7/08C07K16/18C12Q1/68A61K48/00A61K38/17A61K31/7088A61P35/02
Inventor 王少元李景岗黄源茂许能文叶美玲
Owner FUJIAN MEDICAL UNIV UNION HOSPITAL
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