EXPRESSION OF GRANULOCYTE COLONY STIMULATING FACTOR (GCSF) IN HANSENULA POLYMORPHA

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.

TALEBKHAN YEGANEH; SAMADI TANNAZ; SAMIE ARMIN; BARKHORDARI FARZANEH; AZIZI MOHAMMAD; KHALAJ VAHID; MIRABZADEH ESMAT

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

Journal Paper

Paper Information

Journal: Iranian Journal of Microbiology Year:2016 | Volume:8 | Issue:1 Page(s): 21-28

Download Full-Text

View:

260

Download:

Cites:

Information Journal Paper

Title

EXPRESSION OF GRANULOCYTE COLONY STIMULATING FACTOR (GCSF) IN HANSENULA POLYMORPHA

Author(s)

Keywords

HANSENULA POLYMORPHA

EXPRESSION CASSETTE

GCSF

Abstract

Background and Objectives: During past decades HANSENULA POLYMORPHA has attracted global attention for the expression of recombinant proteins due to its high growth rate, minimal nutritional porequirements and use of methanol as a low cost inducer.Materials and Methods: The corresponding nucleotide sequences for the expression of heterologous genes in Hansenula poylmorpha were extracted and assembled in an E. coli vector. The constructed EXPRESSION CASSETTE included formate dehydrogenase promoter (pFMD), a secretory signal sequence, a multiple cloning site (MCS) and methanol oxidase (MOX) terminator. Zeocin resistance gene fragment and complete cDNA encoding granulocyte colony stimulating factor (GCSF) were cloned downstream of the EXPRESSION CASSETTE in-frame with signal sequence. Restriction mapping and sequence analysis confirmed the correct cloning procedures. Final vector was transformed into Hansenula and recombinant host was induced for the expression of GCSF protein by adding methanol. SDS-PAGE and immuno-blotting were performed to confirm the identity of r-GCSF.Results: The EXPRESSION CASSETTE containing GCSF gene (615bp) and zeocin resistance marker (sh-ble, 1200bp) was prepared and successfully transformed into competent HANSENULA POLYMORPHA cells via electroporation. Zeocin resistant colonies were selected and GCSF expression was induced in recombinant Hansenula transformants using 0.5% methanol and an approximately 19kDa protein was observed on SDS-PAGE. Western blot analysis using serum isolated from GCSF-treated rabbit confirmed the identity of the protein.Conclusions: Molecular studies confirmed the designed EXPRESSION CASSETTE containing GCSF gene along with pFMD and signal sequence. The expressed 19kDa protein also confirmed the ability of designed vector in expressing heterologous genes in Hansenula cells.

Cites

No record.

References

No record.

Cite

APA: Copy

TALEBKHAN, YEGANEH, SAMADI, TANNAZ, SAMIE, ARMIN, BARKHORDARI, FARZANEH, AZIZI, MOHAMMAD, KHALAJ, VAHID, & MIRABZADEH, ESMAT. (2016). EXPRESSION OF GRANULOCYTE COLONY STIMULATING FACTOR (GCSF) IN HANSENULA POLYMORPHA. IRANIAN JOURNAL OF MICROBIOLOGY, 8(1), 21-28. SID. https://sid.ir/paper/315388/en

Vancouver: Copy

TALEBKHAN YEGANEH, SAMADI TANNAZ, SAMIE ARMIN, BARKHORDARI FARZANEH, AZIZI MOHAMMAD, KHALAJ VAHID, MIRABZADEH ESMAT. EXPRESSION OF GRANULOCYTE COLONY STIMULATING FACTOR (GCSF) IN HANSENULA POLYMORPHA. IRANIAN JOURNAL OF MICROBIOLOGY[Internet]. 2016;8(1):21-28. Available from: https://sid.ir/paper/315388/en

IEEE: Copy

YEGANEH TALEBKHAN, TANNAZ SAMADI, ARMIN SAMIE, FARZANEH BARKHORDARI, MOHAMMAD AZIZI, VAHID KHALAJ, and ESMAT MIRABZADEH, “EXPRESSION OF GRANULOCYTE COLONY STIMULATING FACTOR (GCSF) IN HANSENULA POLYMORPHA,” IRANIAN JOURNAL OF MICROBIOLOGY, vol. 8, no. 1, pp. 21–28, 2016, [Online]. Available: https://sid.ir/paper/315388/en

Related Journal Papers

No record.

Related Seminar Papers

No record.

Related Plans

No record.

Recommended Workshops