Your browser doesn't support javascript.
loading
Preparation of fusion protein between ZZ affinity peptide and alkaline phosphatase and its primary application / 中国免疫学杂志
Chinese Journal of Immunology ; (12): 1214-1216,1220, 2015.
Article in Chinese | WPRIM | ID: wpr-602259
ABSTRACT

Objective:

To construct and express the fusion protein between ZZ affinity peptide and alkaline phosphatase and examine its biological activities.

Methods:

The alkaline phosphatase gene was cloned into pEZZ 18 vector containing ZZ peptide gene resulted in the pEZZ-AP recombinant vector.Then the vector was transformed and expressed in E.coli DH5α.And HisTrap affinity chromatography was employed to separate and purify the target protein.After analyzed by Western blot , ZZ-AP fusion protein was applied to immunocytochemistry as an alterative second antibody.

Results:

The result of SDS-PAGE showed that the fusion protein with a relative molecular weight was consistent with the theoretical values (62 kD).The fusion protein has rabbit IgG-binding ability and en-zymatic activity of alkaline phosphatase ,those were validated in Western blot;and it produced a good signal that was comparable in its staining pattern to that generated with goat anti-rabbit IgG-AP in immunocytochemistry.

Conclusion:

The ZZ-AP fusion protein was constructed successfully ,it has rabbit IgG-binding ability and enzymatic activity of alkaline phosphatase.We anticipate that the ZZ-AP fusion protein has a potential application in immunoassay field.

Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Immunology Year: 2015 Type: Article

Similar

MEDLINE

...
LILACS

LIS

Full text: Available Index: WPRIM (Western Pacific) Language: Chinese Journal: Chinese Journal of Immunology Year: 2015 Type: Article