[Cloning, prokaryotic expression of CML66 and preparation of its polyclonal antibody].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
; 19(6): 582-4, 2003 Nov.
Article
em Zh
| MEDLINE
| ID: mdl-15182656
ABSTRACT
AIM:
To clone and express CML66 cDNA and to prepare rabbit anti-CML66 antibody.METHODS:
cDNA isolated from the testis using RT-PCR was cloned into pGEMT. After sequencing, the cDNA was inserted into prokaryotic expression vector pET32b(+). The recombinant vector was transformed into BL-21(DE3) through electroporation. 6xHis-tagged CML66 expression was then induced by IPTG. The protein was purified through Ni(2+) affinity chromatography column and characterized by SDS-PAGE and Western blot. The purified protein was injected into rabbits to prepare polyclonal antibody.RESULTS:
The cloned cDNA sequence was identical with that previously reported. The target protein was successfully purified. And rabbit's anti-serum with high titer was obtained.CONCLUSION:
We have cloned CML66 successfully, expressed and purified the protein in E.coli.Furthermore,rabbit polyclonal antibody has been obtained.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Anticorpos
/
Antígenos de Neoplasias
Limite:
Humans
/
Male
Idioma:
Zh
Ano de publicação:
2003
Tipo de documento:
Article