INVESTIGATION OF REASONS THAT IMPLY THE DIMINISHED INCLUSION BODIES IN E. COLI

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MIRZAHOSSEINI H.; OMUMI A.; OMIDINIA E.

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Journal Paper

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Journal: Journal of Sciences, Islamic Republic of Iran Year:2003 | Volume:14 | Issue:2 Page(s): 0-0

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Title

INVESTIGATION OF REASONS THAT IMPLY THE DIMINISHED INCLUSION BODIES IN E. COLI

Author(s)

MIRZAHOSSEINI H. | OMUMI A. | OMIDINIA E. | Issue Writer Certificate

Keywords

INCLUSION BODIES; HBFGF; GROWTH INHIBITION

Abstract

We previously reported the cloning and expression of cDNA of Human Basic Fibroblast Growth Factor (hbFGF) in Escherichia coli under the control of T7 promoter [6]. In this study we looked for the factor which affects the diminution of Inclusion Bodies (IBs) in E. coli cells which produce heterologous proteins (hbFGF), 6 h after induction. It was shown that the amount of the insoluble hbFGF (as inclusion bodies), reached to the highest level 3 h after induction at 37°C and 42°C, however it decreased gradually afterwards to the lowest level at 6 hrs after induction. To address whether this diminution is the result of destruction of IBs by E. coli proteases, cell death, cell growth inhibition or changing in other physiological characteristics of E. coli, we carried out three experiments. These experiments were continual removal of the E. coli cells which loose their plasmids in culture by adding ampicillin; comparing the number of colonies which loose their plasmids to those which keep the plasmids by growing cells on LB agar plates containing ampicillin or without ampicillin; and adding chloramphenicol to culture 3 h after induction in order to cease protein synthesis in the E. coli cells. The results showed that neither IBs destruction by proteases nor cell death caused the IBs diminution. We showed that the IBs reduction is the immediate result of overgrowth of plasmid-free cells in culture and inhibition of proliferation of the cells containing plasmids and producing the heterologous proteins. Consequently, the ratio of heterologous protein producing cells to the ones lack this character is decreased.

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APA: Copy

MIRZAHOSSEINI, H., OMUMI, A., & OMIDINIA, E.. (2003). INVESTIGATION OF REASONS THAT IMPLY THE DIMINISHED INCLUSION BODIES IN E. COLI. JOURNAL OF SCIENCES ISLAMIC REPUBLIC OF IRAN, 14(2), 0-0. SID. https://sid.ir/paper/529072/en

Vancouver: Copy

MIRZAHOSSEINI H., OMUMI A., OMIDINIA E.. INVESTIGATION OF REASONS THAT IMPLY THE DIMINISHED INCLUSION BODIES IN E. COLI. JOURNAL OF SCIENCES ISLAMIC REPUBLIC OF IRAN[Internet]. 2003;14(2):0-0. Available from: https://sid.ir/paper/529072/en

IEEE: Copy

H. MIRZAHOSSEINI, A. OMUMI, and E. OMIDINIA, “INVESTIGATION OF REASONS THAT IMPLY THE DIMINISHED INCLUSION BODIES IN E. COLI,” JOURNAL OF SCIENCES ISLAMIC REPUBLIC OF IRAN, vol. 14, no. 2, pp. 0–0, 2003, [Online]. Available: https://sid.ir/paper/529072/en

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