Prokaryotic expression and purification of SPAG4L, a novel human testis gene / 南方医科大学学报
Journal of Southern Medical University
; (12): 2047-2050, 2010.
Article
in Zh
| WPRIM
| ID: wpr-330787
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To express SPAG4L, a novel human testis gene in E. coli and purify it's fusion protein.</p><p><b>METHODS</b>The fragment encoding SPAG4L126-379 was amplified by RT-PCR and the PCR products were cloned into PUCm-T vectors. After digestion by EcoR I and Hind III, the fragment was subcloned into PQE-30, a prokaryotic expression vector with 6×His tag. The recombinant plasmid PQE-30-SPAG4L was sequenced and transformed into E.coli M15. The expression of his-tagged fusion protein was induced by IPTG. The fusion protein was identified by Western blotting and purified using Ni-NTA magnetic agarose beads.</p><p><b>RESULTS</b>The recombinant plasmid PQE-30-SPAG4L was constructed successfully and expressed in E.coli M15. The fusion protein SPAG4Lwith 6×his-tag was confirmed by Western blotting. The micro-scale purification system of 6×His-tagged SPAG4Lprotein was established and purified fusion protein was obtained.</p><p><b>CONCLUSION</b>The recombinant plasmid PQE-30-SPAG4L can be expressed in vitro and used for studying the biological function of SPAG4L in spermatogenesis.</p>
Full text:
1
Index:
WPRIM
Main subject:
Plasmids
/
Recombinant Fusion Proteins
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Carrier Proteins
/
Escherichia coli
/
Genetics
/
Metabolism
Limits:
Humans
/
Male
Language:
Zh
Journal:
Journal of Southern Medical University
Year:
2010
Type:
Article