CLONING AND EXPRESSION OF RECOMBINANT TISSUE FACTOR IN CHO CELLS

Q: How can I download an article?

To download an article from SID, first log in to the site, search for the article title, and click on the 'Download Article' option.

Q: How can I download an ISI article?

To download an ISI article on SID, enter the keyword or article title in the search bar, view the relevant results, click on the desired article, and select the 'Download Article' option.

Q: How can I access the SID database?

To access the SID database, visit SID.ir, create an account, and log in to access scientific resources.

Q: Is downloading articles from SID free?

Some articles on SID are available for free, while others require payment. Details are specified on the article's page.

KHORSHIDFAR M.; AMIRIZADEH N.; HABIBI ROUDKENAR M.

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Journal Paper

Paper Information

Journal: The Scientific Journal of Iranian Blood Transfusion Year:2011 | Volume:8 | Issue:2 (31) Page(s): 96-103

Download Full-Text

Persian Verion

View:

1,131

Download:

Cites:

Information Journal Paper

Title

CLONING AND EXPRESSION OF RECOMBINANT TISSUE FACTOR IN CHO CELLS

Author(s)

KHORSHIDFAR M. | AMIRIZADEH N. | HABIBI ROUDKENAR M. | Issue Writer Certificate

Keywords

CHO CELLSQ3

FACTOR VIIQ3

PROTHROMBIN TIMEQ3

Abstract

Background and Objectives: Tissue factor (TF), a 45-kDa trans membrane glycoprotein, is the major cellular initiator of the coagulation cascade. PROTHROMBIN TIME (PT) is the test that evaluates extrinsic pathway of coagulation. Thromboplastin used in this test is mostly prepared of rabbit brain. Thus it causes variation in the PT results. There is an important advantage of using recombinant TF in PT test to get more reproducible results.Materials and Methods: TF mRNA was isolated from human lung fibroblast cells. After preparation of cDNA it has cloned in pcDNA3 plasmid. CHO CELLS were transfected with recombinant plasmid.Transfected cells were grown in presence of Geneticin. Total proteins were extracted from CHO CELLS and separated with SDS/PAGE electrophoresis and analyzed by western blotting technique. CHO CELLS expressing TF were added to citrated plasma in presence of CaCl2 and clotting time was measured. In addition, FACTOR VII activation by recombinant TF in the plasma was assessed by ELISA method.Results: Extracted proteins from tarnsfected CHO CELLS separated on SDS/PAGE showed an approximately 40-kDa band that verified with a monoclonal antibody against TF on western blot analysis. Adding transfected cells (106cell/ml) to the citrated plasma in presence of CaCl2 could decrease clotting time from 3 minutes to 21±3 seconds compared with untransfected CHO CELLS. The level of FACTOR VIIa in citrated plasma was 810±35 ng/ml in presence of transfected cells’ membrane.Conclusions: Recombinant TF expressed in CHO CELLS was able to clot citrated plasma and to activate FACTOR VII.

Cites

No record.

References

No record.

Cite

APA: Copy

KHORSHIDFAR, M., AMIRIZADEH, N., & HABIBI ROUDKENAR, M.. (2011). CLONING AND EXPRESSION OF RECOMBINANT TISSUE FACTOR IN CHO CELLS. THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON), 8(2 (31)), 96-103. SID. https://sid.ir/paper/78512/en

Vancouver: Copy

KHORSHIDFAR M., AMIRIZADEH N., HABIBI ROUDKENAR M.. CLONING AND EXPRESSION OF RECOMBINANT TISSUE FACTOR IN CHO CELLS. THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON)[Internet]. 2011;8(2 (31)):96-103. Available from: https://sid.ir/paper/78512/en

IEEE: Copy

M. KHORSHIDFAR, N. AMIRIZADEH, and M. HABIBI ROUDKENAR, “CLONING AND EXPRESSION OF RECOMBINANT TISSUE FACTOR IN CHO CELLS,” THE SCIENTIFIC JOURNAL OF IRANIAN BLOOD TRANSFUSION ORGANIZATION (KHOON), vol. 8, no. 2 (31), pp. 96–103, 2011, [Online]. Available: https://sid.ir/paper/78512/en

Related Journal Papers