Expression and biology identification of the human epididymis-specific gene ESC42 in E. coli / 中华男科学杂志
National Journal of Andrology
;
(12): 106-111, 2005.
Article
in Chinese
| WPRIM
| ID: wpr-267744
ABSTRACT
<p><b>OBJECTIVE</b>To provide materials for the study of the function of ESC42 protein specifically expressed in the human epididymis.</p><p><b>METHODS</b>The ESC42 gene was amplified from the human epididymis cDNA library by PCR and then cloned into prokaryotic expression vector pGEX-4T-1, expressed and purified by recombinant DNA techniques. The specificity of ESC42 protein was identified by Western blot and MALDI-TOF-MS. The database was searched by Ms-Fit.</p><p><b>RESULTS</b>The recombinant plasmid expressed a Mr 38 x 10(3) fusion protein in E. coli at a level of 30% of the total protein, and the purity was as high as 99%. The ESC42 protein was identified by ESC42 monoclonal antibody and its molecular weight was 11 978.12, tested by MALDI-TOF-MS. The peptide mass fingerprint analysis showed that the coverage rate of the sequence reached 48% with 100% matching. The motif scan in Prosite database reveal that ESC42 belonged to the beta-defensin family and had antibacterial activity.</p><p><b>CONCLUSION</b>Obtaining high purity of rhESC42 protein may lay a foundation for the study of its functions.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Recombinant Fusion Proteins
/
Gene Library
/
Amino Acid Sequence
/
Cloning, Molecular
/
Defensins
/
Allergy and Immunology
/
Epididymis
/
Escherichia coli
/
Genetics
Type of study:
Diagnostic study
Limits:
Animals
/
Humans
/
Male
Language:
Chinese
Journal:
National Journal of Andrology
Year:
2005
Type:
Article
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