Construction and identification of recombinant adenoviral vector encoding B7-H4 gene / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
; (6): 499-505, 2008.
Artigo
em Chinês
| WPRIM (Pacífico Ocidental)
| ID: wpr-310423
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To construct the recombinant adenovirus containing B7-H4 gene with AdEasy XL system and to identify its biological activities.</p><p><b>METHODS</b>The full-length mouse B7-H4 gene was amplified by RT-PCR from C57 mouse lung and put into T vector, then verified by sequencing. Digested with Xhol I and EcoR V the B7-H4 gene was inserted into pshuttle-CMW(PSC). Pme I linearized shuttle plasmid was transformed into E.coli BJ5183-AD-1 to obtain the recombinant adenoviral plasmid pAd-mB7-H4 by efficient homologous recombination. Then the recombinant adenovirus-mB7-H4/Ad was obtained by packaging Pac I linearized in D-293 cells. The mRNA and protein expression of B7-H4 in mB7-H4/Ad infected AD-293 cells were detected by RT-PCR and Western blot, respectively. mB7-H4/Ad was used to infect L929 cells, the bioactivity of expressed B7-H4 in stimulation of T lymphocytes proliferation and cytokine production were tested.</p><p><b>RESULTS</b>The full-length of mB7-H4 was cloned from mouse lung tissue cDNA and verified by sequencing. The recombinant plasmid pAd-m B7-H4 was successfully generated by homologous recombination, and the primary mB7-H4/Ad was obtained by packaging pAd-B7-H4 in AD-293 cells. Compared with the negative control, L929 cells infected with mB7-H4/Ad effectively inhibited the proliferation of T lymphocytes and cytokines production.</p><p><b>CONCLUSION</b>The bioactive recombinant adenovirus mB7-H4/Ad has been successfully constructed.</p>
Texto completo:
Disponível
Contexto em Saúde:
Doenças Negligenciadas
Problema de saúde:
Doenças Negligenciadas
/
Zoonoses
Base de dados:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Proteínas Recombinantes de Fusão
/
Linfócitos T
/
Transfecção
/
Adenoviridae
/
Clonagem Molecular
/
Antígeno B7-1
/
Alergia e Imunologia
/
Escherichia coli
/
Inibidor 1 da Ativação de Células T com Domínio V-Set
Limite:
Animais
Idioma:
Chinês
Revista:
Journal of Zhejiang University. Medical sciences
Ano de publicação:
2008
Tipo de documento:
Artigo