[The expression of C-type lectin-like domain of DNGR-1 molecules of Chinese rhesus macaques in E.coli].

Autor:	Yao W; National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China., Yang G; National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Jazyk:	čínština
Zdroj:	Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology [Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi] 2014 May; Vol. 30 (5), pp. 489-92.
Abstrakt:	Objective: To obtain the nucleotide sequence and express the C-type lectin-like domain (CTLD) of C-type lectin domain family 9, member A (Clec9A) gene of Chinese rhesus macaques. Methods: The complete coding region of rhesus macaque Clec9A gene was amplified by sequence specific RT-PCR and sub-cloned into the pGEM-T vector and sequenced. Recombinant plasmid pET-32a-CTLD was constructed to express the CTLD region of CLEC9A in BL21 (DE3) cells under IPTG induction. The recombinant protein was identified by SDS-PAGE and Western blotting. Results: The 726 bp complete coding sequence of Clec9A gene was obtained. Sequencing and sequence alignments showed that rhesus macaque Clec9A gene shared 95.0% and 73.1% nucleotide identities and 91.7% and 57.3% amino acid identities with humans and mice, respectively. In vitro expression of CTLD resulted in a protein with Mr; of 32 000. Western blotting revealed that the protein had the similar antigenicity with human DNGR-1. Conclusion: The Clec9A gene of Chinese rhesus macaques is highly similar to that of humans and could code a protein with the antigenicity of human CTLD protein.
Databáze:	MEDLINE
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