COMPARISON OF DIFFERENT METHODS OF OSMOTIC SHOCKS FOR EXTRACTION OF HUMAN GRANULOCYTE COLONY STIMULATING FACTOR PRODUCED IN PERIPLASM

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.

PEYMANFAR SHARAREH; ROGHANIAN RASOUL; GHAEDI KAMRAN

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

Journal Paper

Paper Information

Journal: BIOLOGICAL JOURNAL OF MICROORGANISM Year:2018 | Volume:7 | Issue:26 Page(s): 43-50

Download Full-Text

Persian Verion

View:

1,035

Download:

Cites:

Information Journal Paper

Title

COMPARISON OF DIFFERENT METHODS OF OSMOTIC SHOCKS FOR EXTRACTION OF HUMAN GRANULOCYTE COLONY STIMULATING FACTOR PRODUCED IN PERIPLASM

Author(s)

PEYMANFAR SHARAREH | ROGHANIAN RASOUL | GHAEDI KAMRAN | Issue Writer Certificate

Keywords

E. COLIQ2

G-CSFQ1

PET22BQ2

INTEINQ1

PELBQ1

OSMOTIC SHOCKQ1

Abstract

Introduction: In the recent years, a growing interest for the production of secretary recombinant proteins is seen. This is due to the advantages of recombinant protein production in the periplasm compared to the cytoplasm. However, signal peptides have a critical role in protein secretion as well as the applied technique for the extraction of the protein. Granulocyte colony stimulating factor (GCSF) is a type of colony stimulating factor that causes motivating of proliferation, differentiation and survival of neutrophiles and progenitor cells of these cells and are used to promote decreased neutrophils in some cancers after chemotherapy. The aim of this study was to evaluate the usage of different OSMOTIC SHOCK assays in order to achieve the highest amount of granulocyte colony stimulating factor (GCSF) in BL21 strain of E. COLI.Materials and methods: The E. COLI which contained PET22B- GCSF2- INTEIN2 expression vector was cultured in 4YT medium and was induced with IPTG 1Mm for protein production. It is necessary to mention that the PET22B has a PELB signal peptide that directs proteins to the periplasmic space. In the next step, three different methods of OSMOTIC SHOCKs were applied for the extraction of the obtained human recombinant protein. Finally, the isolated proteins were analyzed by SDS-PAGE and western blot techniques.Results: The results of this investigation indicated that GCSF was produced in both of the cytoplasmic and periplasmic spaces and the best method of OSMOTIC SHOCK for protein extraction is using Tris buffer and MgSO4.Discussion and conclusion: Regarding the results, it is concluded that the MgSO4 with Tris buffer create a good osmotic pressure and accordingly is a more effective way for G-CSF protein extraction. As a result, this method could be used for production and simple separation of recombinant drug proteins.

Cites

Characterization and In Silico Analysis of The Structural Features of G-CSF Derived from Lysates of Escherichia coli

References

No record.

Cite

APA: Copy

PEYMANFAR, SHARAREH, ROGHANIAN, RASOUL, & GHAEDI, KAMRAN. (2018). COMPARISON OF DIFFERENT METHODS OF OSMOTIC SHOCKS FOR EXTRACTION OF HUMAN GRANULOCYTE COLONY STIMULATING FACTOR PRODUCED IN PERIPLASM. BIOLOGICAL JOURNAL OF MICROORGANISM, 7(26 ), 43-50. SID. https://sid.ir/paper/237485/en

Vancouver: Copy

PEYMANFAR SHARAREH, ROGHANIAN RASOUL, GHAEDI KAMRAN. COMPARISON OF DIFFERENT METHODS OF OSMOTIC SHOCKS FOR EXTRACTION OF HUMAN GRANULOCYTE COLONY STIMULATING FACTOR PRODUCED IN PERIPLASM. BIOLOGICAL JOURNAL OF MICROORGANISM[Internet]. 2018;7(26 ):43-50. Available from: https://sid.ir/paper/237485/en

IEEE: Copy

SHARAREH PEYMANFAR, RASOUL ROGHANIAN, and KAMRAN GHAEDI, “COMPARISON OF DIFFERENT METHODS OF OSMOTIC SHOCKS FOR EXTRACTION OF HUMAN GRANULOCYTE COLONY STIMULATING FACTOR PRODUCED IN PERIPLASM,” BIOLOGICAL JOURNAL OF MICROORGANISM, vol. 7, no. 26 , pp. 43–50, 2018, [Online]. Available: https://sid.ir/paper/237485/en

Related Journal Papers