[Construction of a prokaryotic expression vector for apoptin and preparation of polyclonal antibody of apoptin].
OBJECTIVE
To construct a prokaryotic expression vector for apoptin and prepare polyclonal antibody of apoptin.


METHODS
Apoptin gene amplified from pGEM-T/Apoptin plasmid by PCR was cloned into pET-28a (+). E.coli BL21 (DE3) was transformed by the recombinant plasmid, and apoptin protein expression induced by IPTG was analyzed by SDS-PAGE. BALB/c mice were immunized with the protein and the titer of the antibody was determined using indirect enzyme-linked immunosorbent assay (ELISA).


RESULTS
Apoptin gene was successfully cloned into pET-28a (+), and the expression of a protein with relative molecular mass of about 17 000 was identified by SDA-PAGE. After 5 immunizations of the mice with the protein, the blood antibody titer reached 1:5x10(5).


CONCLUSIONS
The prokaryotic expression vector for apoptin is successfully constructed and the polyclonal antibody of apoptin is obtained, which allows further functional study of apoptin.
Wang
Jian-Sheng
JS
Department of Oncological Surgery, First Affiliated Hospital, Xi'an Jiaotong University College of Medicine, Xi'an 710061, China. wangjsh@mail.xjtu.edu.cn
Zhang
Ming-Xin
MX
Duan
Xiao-Yi
XY
Mo
Fei
F
Wang
Quan-Ying
QY
Yang
Guang-Xiao
GX
D000818:Animals; D000906:Antibodies / Q000097:blood* / Q000235:genetics / Q000276:immunology; D036022:Capsid Proteins / Q000235:genetics* / Q000276:immunology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D016678:Genome; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids
D000906:Antibodies; D036022:Capsid Proteins; C091717:VP3 protein, Chicken anemia virus