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Two kinds of novel upexpressed human protein in liver cancer and its coding sequence, and novel uses of other twenty kinds of human protein in liver cancer diagnosis

A human protein and sequence technology, applied in the use and preparation of polynucleotides and polypeptides, polypeptides encoded by polynucleotides, and the diagnosis of liver cancer, can solve the problem of lack of high-throughput functional genomes

Inactive Publication Date: 2006-05-03
上海博星基因芯片有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Human genomics research is currently a hot spot in the world. In addition to large-scale sequencing of human chromosome DNA and expressed sequence sequencing (EST), there is still a lack of high-throughput methods for screening functional genomes starting from functions.

Method used

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  • Two kinds of novel upexpressed human protein in liver cancer and its coding sequence, and novel uses of other twenty kinds of human protein in liver cancer diagnosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0116] Example 1: Cloning of Human Protein Upregulated in Liver Cancer Tissue

[0117] 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 Qiagene). 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 Dye terminate 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 sequences of two clones, PP0174B04 and PP1277E03, were new DNA. The insert cDNA fragment contained ...

Embodiment 2

[0118] Example 2: Homology retrieval of cDNA clones

[0119] The two new human proteins and their coding sequences up-regulated in liver cancer tissues of the present invention were used to Blast program (Basiclocal Alignment search tool) [Altschul, SF et al.J.Mol.Biol.1990; 215:403- 10], and performed homology searches in databases such as Genbank and Swissport. The protein homology results are as follows:

[0120] 1. PP0174B04 protein

[0121] Blastp

[0122] Query=PP0174B04 (162 amino acids)

[0123] >gi|34532417|dbj|BAC86421.1|unnamed protein product[Homo sapiens]

[0124] gi|46409598|ref|NP_997369.1|FLJ44076 protein[Homo sapiens]

[0125] length = 181

[0126] Score = 53.5bits (127), predicted value = 2e-06

[0127] Identity = 31 / 72 (43%), Similarity = 34 / 72 (47%), Gap = 1 / 72 (1%)

[0128] Query: 65 TFHCKHLLATPAVAQAGPGVAHDTILEVTNCKLWWHLHGTNSAGTQNVRTVETCLPSPRP 124

[0129] T C + PAVAQ GPG A T E N K WW H G Q+ R VE P P

[0130] Sbjct: 111 ...

Embodiment 3

[0159] Example 3: Study on Expression Profile of Hepatocellular Carcinoma (HCC) Using cDNA Chip

[0160]Gene chip or gene microarray (DNA Microarray) is a new technology that many national laboratories and large pharmaceutical companies are currently developing and developing. It refers to the orderly and high-density arrangement of a large number of target gene fragments on glass, Then use fluorescence detection and computer software to compare and analyze the data, so as to achieve the purpose of rapid, efficient and high-throughput analysis of biological information. The polynucleotide of the present invention can be used as target DNA in gene chip technology for high-throughput research on new gene functions; finding and screening tissue-specific new genes, especially new genes related to diseases such as tumors; disease diagnosis, such as genetic diseases . Its specific method steps have multiple reports in the literature, as can refer to the literature DeRisi, J.L., Lye...

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Abstract

The invention discloses two human albumens expressed in liver cancer group, polynucleotide to code the polypeptide, generation method for the polypeptide with DNA recombination technique, application method of this polypeptide for cancer, the v to this polypeptide, and other twenty human albumens in liver cancer group with their codes and new application (there was no opposite report).

Description

Technical field: [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to two new polynucleotides encoding human proteins expressed up-regulated in liver cancer tissues, and polypeptides encoded by the polynucleotides. The present invention also relates to the use and preparation of such polynucleotides and polypeptides. The present invention also relates to the novel use of the other twenty human proteins and their coding sequences up-regulated in liver cancer tissue in the diagnosis of liver cancer (no one has reported its use in the diagnosis of liver cancer before). Background technique: [0002] Human genomics research is currently a hot spot in the world. In addition to large-scale sequencing of human chromosomal DNA and expressed sequence sequencing (EST), there is still a lack of high-throughput methods for screening functional genomes starting from functions. [0003] Cancer is one of the major diseases that endanger human...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K14/47C12N15/12C12N15/63C12Q1/68
Inventor 孙美倩李瑶裘敏燕谢毅毛裕民
Owner 上海博星基因芯片有限责任公司
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