Construction of mammalian cell expression vector of human CD154 gene from active peripheral blood mononuclear cell and analysis of its sequence / 免疫学杂志
Immunological Journal
;
(12): 88-90, 2001.
Artigo
em Chinês
| WPRIM
| ID: wpr-433878
ABSTRACT
Objective To obtain mammalian cell expression vector of human CD154 gene. Methods A 820 bp cDNA fragment was amplified by RT-PCR method from total RNA of human peripheral blood mononuclear cell(PBMC) activated with 10 ng/mL PMA and 1 μg/mL PHA for 8 hours. The fragment was cloned into pcDNA3.1(+) plasmids.The cloned insert was identified by double digestion of the recombinant plasmid with restriction enzymes BamH Ⅰ and EcoR Ⅰ and sequenced by Sangers-dideory-mediated chain termination. Results This cDNA fragment included 820 bp entire coding region and a part of the 3 non-coding region. The recombinant mammalian cell expression vector of pcDNA3.1(+)/hCD154 was constructed, the sequence of the insert was identical to the published sequence encoding human CD154 antigen. Conclusion The recombinant mammalian cell expression vector of pcDNA3.1(+)/hCD154 was successfully constructed.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
Immunological Journal
Ano de publicação:
2001
Tipo de documento:
Artigo
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