TRANSFORMATION OF CHOLERA TOXIN B SUBUNIT-HUMAN PROINSULIN CHIMERIC GENE TO EDIBLE MUSHROOM (PLEUROTUS OSTREATUS) AND EVALUATION OF ITS PRESENCE AND EXPRESSION

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FABRIKI OURANG S.; JALALI JAVARAN M.; MOHAMMADI GOLTAPEH E.; ALIZADEH H.; HONARI H.

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Journal: MODERN GENETICS JOURNAL (MGJ) Year:2017 | Volume:12 | Issue:3 Page(s): 441-449

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Title

TRANSFORMATION OF CHOLERA TOXIN B SUBUNIT-HUMAN PROINSULIN CHIMERIC GENE TO EDIBLE MUSHROOM (PLEUROTUS OSTREATUS) AND EVALUATION OF ITS PRESENCE AND EXPRESSION

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Abstract

Diabetes is a disease in which the body does not produce or properly utilize insulin. Although insulin is predominantly made in bacteria or yeast for commercial production, there is a burgeoning need for alternative systems in the production of insulin. In this regard, edible oyster mushroom was studied as host for the production of insulin. Therefore, a binary vector pCAMBIACTB-INS was constructed containing the cholera toxin B subunit fused with human proinsulin gene under the control of glyceraldehyde-3-phosphate dehydrogenase as homologous promoter. Then it introduced into gill tissue of edible oyster mushroom, Pleurotus ostreatus isolate Iran1649c, via Agrobacterium Tumefaciens strain GV3101 (OD600nm) with co-cultivation at 25oC and selective medium containing hygromycin and cefotaxime. After confirming Putative transformants using PCR with specific primers, stable integration of the CTB-proinsulin fusion gene into P. ostreatus genome was confirmed by Southern blotting analysis. Also, the expression of transgene was confirmed by RT-PCR, ELISA and immunoblotting methods and in transgenic mycelia the recombinant CTB-proinsulin showed an expression level by 0.04% of total soluble protein. In conclusion successful expression of foreign genes in transgenic mushrooms is promising for development of biopharmaceutical in edible mushrooms.

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APA: Copy

FABRIKI OURANG, S., JALALI JAVARAN, M., MOHAMMADI GOLTAPEH, E., ALIZADEH, H., & HONARI, H.. (2017). TRANSFORMATION OF CHOLERA TOXIN B SUBUNIT-HUMAN PROINSULIN CHIMERIC GENE TO EDIBLE MUSHROOM (PLEUROTUS OSTREATUS) AND EVALUATION OF ITS PRESENCE AND EXPRESSION. MODERN GENETICS JOURNAL (MGJ), 12(3 ), 441-449. SID. https://sid.ir/paper/116521/en

Vancouver: Copy

FABRIKI OURANG S., JALALI JAVARAN M., MOHAMMADI GOLTAPEH E., ALIZADEH H., HONARI H.. TRANSFORMATION OF CHOLERA TOXIN B SUBUNIT-HUMAN PROINSULIN CHIMERIC GENE TO EDIBLE MUSHROOM (PLEUROTUS OSTREATUS) AND EVALUATION OF ITS PRESENCE AND EXPRESSION. MODERN GENETICS JOURNAL (MGJ)[Internet]. 2017;12(3 ):441-449. Available from: https://sid.ir/paper/116521/en

IEEE: Copy

S. FABRIKI OURANG, M. JALALI JAVARAN, E. MOHAMMADI GOLTAPEH, H. ALIZADEH, and H. HONARI, “TRANSFORMATION OF CHOLERA TOXIN B SUBUNIT-HUMAN PROINSULIN CHIMERIC GENE TO EDIBLE MUSHROOM (PLEUROTUS OSTREATUS) AND EVALUATION OF ITS PRESENCE AND EXPRESSION,” MODERN GENETICS JOURNAL (MGJ), vol. 12, no. 3 , pp. 441–449, 2017, [Online]. Available: https://sid.ir/paper/116521/en

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