THE INCREASE IN PROTEIN AND PLASMID YIELDS OF E. COLI WITH OPTIMIZED CONCENTRATION OF AMPICILLIN AS SELECTION MARKER

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.

FEIZOLLAHZADEH SADEGH; Kouhpayeh Shirin; RAHIMMANESH ILNAZ; KHANAHMAD HOSSEIN; Sabzehei Faezeh; GANJALIKHANI HAKEMI MAZDAK; ANDALIB ALIREZA; HEJAZI ZAHRA; REZAEI ABBAS

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

Journal Paper

Paper Information

Journal: Iranian Journal of Biotechnology Year:2017 | Volume:15 | Issue:2 Page(s): 0-0

Download Full-Text

View:

472

Download:

Cites:

Information Journal Paper

Title

THE INCREASE IN PROTEIN AND PLASMID YIELDS OF E. COLI WITH OPTIMIZED CONCENTRATION OF AMPICILLIN AS SELECTION MARKER

Author(s)

Keywords

AMPICILLIN

ESCHERICHIA COLI

PLASMID

PROTEIN

Abstract

Background: ESCHERICHIA COLI is still the common host for ing and heterologous PROTEIN expression. Various strategies have been employed to increase PROTEIN expression inE. coli, but, it seems that external factors such as selection marker concentration can drastically affect the yield of PROTEIN and PLASMID.Objectives: Alterations of PROTEIN expression and PLASMID yields of E. coli in different concentrations of AMPICILLIN, as selection marker, will be determined. In order to improve heterologous expression, the system will be redesigned and optimized.Materials and Methods: The expression cassette of codon optimized EGFP for E. coli was synthesized in pUC57. The pUC57-GFP was transformed into E. coli Top10F’. The expression of GFP was verified by SDS-PAGE and flow cytometry after induction by IPTG (0.5 mM) and incubation with 0, 100, 200 and 300 μg.mL-1 AMPICILLIN. PLASMID copy numbers of samples were determined by Real-Time PCR on AMP gene using regression line of diluted standard curve.Results: GFP expressing clones formed fair green colonies on LB agar supplemented with 0.5 mM IPTG and showed fluorescence in FL1 filter of flow cytometry and an extra PROTEIN band on SDS-PAGE gel. The fluorescent intensity of GFP in 0, 100, 200 and 300 mg.mL-1 AMPICILLIN in medium were 549.83, 549.78, 1443.52, 684.87, and PLASMID copy numbers were 6.07×109, 3.21×109, 2.32×1010, 8.11×108, respectively. The PLASMID yields were 55 ng.mL-1, 69 ng.μL-1, 164 ng. mL-1 and 41 ng. mL-1, respectively.Conclusion: PROTEIN and PLASMID yields of E. coli are variable in different concentrations of AMPICILLIN and need to be optimized in newly designed expression systems. PROTEIN and PLASMID yield in the optimized concentration (200 mg.mL-1) was signifi cantly (p<0.01) higher than other doses.

Cites

No record.

References

No record.

Cite

APA: Copy

FEIZOLLAHZADEH, SADEGH, Kouhpayeh, Shirin, RAHIMMANESH, ILNAZ, KHANAHMAD, HOSSEIN, Sabzehei, Faezeh, GANJALIKHANI HAKEMI, MAZDAK, ANDALIB, ALIREZA, HEJAZI, ZAHRA, & REZAEI, ABBAS. (2017). THE INCREASE IN PROTEIN AND PLASMID YIELDS OF E. COLI WITH OPTIMIZED CONCENTRATION OF AMPICILLIN AS SELECTION MARKER. IRANIAN JOURNAL OF BIOTECHNOLOGY, 15(2), 0-0. SID. https://sid.ir/paper/695839/en

Vancouver: Copy

FEIZOLLAHZADEH SADEGH, Kouhpayeh Shirin, RAHIMMANESH ILNAZ, KHANAHMAD HOSSEIN, Sabzehei Faezeh, GANJALIKHANI HAKEMI MAZDAK, ANDALIB ALIREZA, HEJAZI ZAHRA, REZAEI ABBAS. THE INCREASE IN PROTEIN AND PLASMID YIELDS OF E. COLI WITH OPTIMIZED CONCENTRATION OF AMPICILLIN AS SELECTION MARKER. IRANIAN JOURNAL OF BIOTECHNOLOGY[Internet]. 2017;15(2):0-0. Available from: https://sid.ir/paper/695839/en

IEEE: Copy

SADEGH FEIZOLLAHZADEH, Shirin Kouhpayeh, ILNAZ RAHIMMANESH, HOSSEIN KHANAHMAD, Faezeh Sabzehei, MAZDAK GANJALIKHANI HAKEMI, ALIREZA ANDALIB, ZAHRA HEJAZI, and ABBAS REZAEI, “THE INCREASE IN PROTEIN AND PLASMID YIELDS OF E. COLI WITH OPTIMIZED CONCENTRATION OF AMPICILLIN AS SELECTION MARKER,” IRANIAN JOURNAL OF BIOTECHNOLOGY, vol. 15, no. 2, pp. 0–0, 2017, [Online]. Available: https://sid.ir/paper/695839/en

Related Journal Papers

No record.

Related Seminar Papers

No record.

Related Plans

No record.

Recommended Workshops