The expression and purificiation of human Annexin Ⅴ overexpressed in E. coli and the detection of apoptosis / 北京大学学报(医学版)
Journal of Peking University(Health Sciences)
;
(6): 167-169, 2001.
Article
in Chinese
| WPRIM
| ID: wpr-411035
ABSTRACT
Objective:
in order to provide rapid and reliable method.Methods:
Encoded Annexin Ⅴ cDNA was amplifyed from U937 cDNA libary by PCR and then subcloned into E coli expression vector. MS2-Annexin Ⅴ fusion protein could be overexpressed in E coli. The MS2 bacteria protein could be removed by thrombin digestion.The mature Annexin Ⅴ was obtained by ion exchange chromatography and the FITC labled Annexin Ⅴ could be used in the detection of apoptosis.Results:
Up to 37% of the total bacterial proteins was rhAnnexin Ⅴ as showed by SDS-PAGE. The purification of Annexin Ⅴ is over 99%. The FITC labled Annexin Ⅴ could efficiently detect apoptosis.Conclusion:
We successfully established the technique procedure of obtaining a large quantity of Annexin Ⅴ and provided the basic routine for popularizing the detection of apoptosis' with high effciency.
Full text:
Available
Index:
WPRIM (Western Pacific)
Type of study:
Diagnostic study
Language:
Chinese
Journal:
Journal of Peking University(Health Sciences)
Year:
2001
Type:
Article
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