BICISTRONIC EXPRESSION OF THE GENES ENCODING THE TRASTUZUMAB MONOCLONAL ANTIBODY IN TOBACCO USING AN INTERNAL RIBOSOME ENTRY SITE SEQUENCE

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JAFARI M.; EHSASATVATAN M.

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Journal: Journal of Molecular and Cellular Researches Year:2016 | Volume:29 | Issue:2 Page(s): 153-169

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Title

BICISTRONIC EXPRESSION OF THE GENES ENCODING THE TRASTUZUMAB MONOCLONAL ANTIBODY IN TOBACCO USING AN INTERNAL RIBOSOME ENTRY SITE SEQUENCE

Author(s)

JAFARI M. | EHSASATVATAN M. | Issue Writer Certificate

Keywords

MONOCLONAL ANTIBODY TRASTUZUMABQ1

BICISTRONIC EXPRESSIONQ1

TOBACCO HEATSHOCK FACTOR-1Q1

INTERNAL RIBOSOMAL ENTRY SITE (IRES)Q1

Abstract

Monoclonal antibodies are heterotetramers consisting of four peptide chains-two identical light (L) chains and two identical heavy (H) chains and simultaneous equimolar expression of the corresponding genes is required to stable produce and assembly into a functional antibody. In order to co-express of multiple genes, different systems have been developed such as internal ribosome entry site (IRES) based vectors. IRES sequences are genetic elements that can drive cap-independent translation initiation of cistrons located downstream of the element. In this study, we constructed a bicsitronic vector for co-expression of genes encoding the H and L chains of the antibreast cancer antibody trastuzumab (Herceptin®) using an IRES sequence derived from the Nicotiana tabacum heat-shock factor 1 (NtHSF1). The effectiveness of construction was evaluated in N. tabacum using agroinfiltration method. Bisictronic expression of transgenes at the RNA transcription level was detected by semi-quantitative RT-PCR analysis. Western blot analyses revealed the efficient accumulation of tetrameric form of trastuzumab in tobacco leaf tissues and antibody content were ca. 0.44% of total soluble protein within 7 days after infiltration. The results of this study demonstrate the efficacy of a plant derived IRES element for BICISTRONIC EXPRESSION of heteromultimeric proteins.

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APA: Copy

JAFARI, M., & EHSASATVATAN, M.. (2016). BICISTRONIC EXPRESSION OF THE GENES ENCODING THE TRASTUZUMAB MONOCLONAL ANTIBODY IN TOBACCO USING AN INTERNAL RIBOSOME ENTRY SITE SEQUENCE. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), 29(2 ), 153-169. SID. https://sid.ir/paper/248520/en

Vancouver: Copy

JAFARI M., EHSASATVATAN M.. BICISTRONIC EXPRESSION OF THE GENES ENCODING THE TRASTUZUMAB MONOCLONAL ANTIBODY IN TOBACCO USING AN INTERNAL RIBOSOME ENTRY SITE SEQUENCE. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY)[Internet]. 2016;29(2 ):153-169. Available from: https://sid.ir/paper/248520/en

IEEE: Copy

M. JAFARI, and M. EHSASATVATAN, “BICISTRONIC EXPRESSION OF THE GENES ENCODING THE TRASTUZUMAB MONOCLONAL ANTIBODY IN TOBACCO USING AN INTERNAL RIBOSOME ENTRY SITE SEQUENCE,” JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), vol. 29, no. 2 , pp. 153–169, 2016, [Online]. Available: https://sid.ir/paper/248520/en

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