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Zone 2 protein of programmed cell death protein 2 analogue and uses thereof

A technology of programmed death, analogs, applied to the protein fragment of programmed death protein analogs - domain 2 protein field

Inactive Publication Date: 2009-01-28
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] So far, there is no report on the function of any fragment protein of programmed death protein 2 analog

Method used

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  • Zone 2 protein of programmed cell death protein 2 analogue and uses thereof
  • Zone 2 protein of programmed cell death protein 2 analogue and uses thereof
  • Zone 2 protein of programmed cell death protein 2 analogue and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The region 2 protein of the programmed death protein 2 analog is prepared by prokaryotic bacterial fusion expression and then cutting out the amino acid sequence of the carrier, and the preparation method includes the following steps:

[0041] (1) RNA was extracted from gastric cancer cell line SGC-7901 by Trizol method using Trizol kit (purchased from Invitrogen Company), and PrimeScript 1 was used to extract RNA st The strand cDNA Synthesis Kit (purchased from Takara Company) synthesized cDNA as a template for PCR by the method of reverse transcription;

[0042] Use Taq enzyme PrimeSTAR HS DNA Polymerase (purchased from Takara Company) to combine the following primers: ATA GATATC ccagaccacaa ctt cc (GATATC is the EcoRV restriction site) and CCG CTC GAG tta cgg tgt atc tgt tac (CTCGAG is the XhoI restriction site) , pull out the nucleic acid sequence of PDCD2like region 2 by the method of PCR amplification; the PCR conditions are:

[0043] 95°C for 3min, 35cycles of ...

Embodiment 2

[0047] The region 2 protein of the programmed death protein 2 analog is prepared by prokaryotic bacterial fusion expression and then cutting out the amino acid sequence of the carrier, and the preparation method includes the following steps:

[0048] (1) Use the RNeasy mini kit purchased from Qiagen to obtain RNA from the liver cancer cell line HepG2, and use PrimeScript 1 st The strand cDNA Synthesis Kit (purchased from Takara Company) synthesized cDNA as a template for PCR by the method of reverse transcription;

[0049] Use Taq enzyme PrimeSTAR HS DNA Polymerase (purchased from Takara Company) to combine the following primers: ATA GATATC ccagaccacaa ctt cc (GATATC is the EcoRV restriction site) and CCG CTC GAG tta cgg tgt atc tgt tac (CTCGAG is the XhoI restriction site) , pull out the nucleic acid sequence of PDCD2like region 2 by the method of PCR amplification; the PCR conditions are:

[0050] 95°C for 3min, 35cycles of 98°C for 15s, 45°C for 30s, 72°C for 1min, the PC...

Embodiment 3

[0054] 98 children with asthma: 35 cases of syncytial virus infection, 19 cases of bacterial (Streptococcus pneumoniae, Moraxella catarrhalis, influenza (Haemophilus) bacillus) infection, 9 cases of mycoplasma infection, 35 cases of unknown etiology, 13 cases of normal children , extract 1 ml of peripheral blood, separate white blood cells with ficoll separation medium, add 1 ml Trizol to extract RNA, use reverse transcriptase to reverse transcribe the RNA into cDNA, and analyze the expression of programmed death protein 2 analog domain 2 by real-time quantitative PCR 18s rRNA was used as internal reference. Base 2, C of 18s rRNA T Value minus programmed death protein 2 analogue region 2 C T The value is an exponent, ie: 2 -(CT，PDCD2like-CT，18s rRNA) , represents the relative expression level of programmed death protein 2 analog domain 2 in each sample.

[0055] from image 3 It can be seen that the expression of programmed death protein 2 analog domain 2 in patients with ...

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Abstract

The invention discloses a protein segment of programmed death protein 2 analog, namely, the protein of region 2, and the application of the protein of the region 2 in preparing drugs for curing bacterial inflammation of the programmed death protein 2 analog. The protein of region 2 of the programmed death protein 2 analog realizes the function of restraining the tumor necrosis factor Alpha of a pro-inflammatory cytokine caused by bacterial LPS, thereby restraining the generating and developing of the inflammations caused by bacteria.

Description

technical field [0001] The present invention relates to a protein fragment of the programmed death protein analogue—area 2 protein, and in particular to the application of the domain 2 protein of the programmed death protein 2 analogue in the preparation of medicines for treating bacterial inflammation. Background technique [0002] Such as figure 1 with figure 2 As shown, programmed death protein 2 analog (PDCD2like, NP_115722) is a similar protein of programmed death protein 2 (PDCD2, NP_002589), which contains an incomplete PDCD2_C domain. [0003] In 1991, Owens G.P. et al first reported programmed death protein 2 (PDCD2), and believed that PDCD2 belongs to a protein related to apoptosis (see: Molecular and Cellular Biology 11, 4177-4188, 1991. Article Identification of mRNAs associated with programmed cell death in immune thymocytes). [0004] But the following two articles suggest the opposite: [0005] Article Cloning of mouse RP-8 cDNA and its expression during ...

Claims

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Application Information

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IPC IPC(8): C07K14/47C12N15/12A61K38/17A61K31/7088A61P31/04A61P31/06
Inventor 陈秋
Owner SUZHOU UNIV
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