EXPRESSION OF RECOMBINANT PROTEIN IN TOBACCO SEEDS

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BAGHERI KH.; GHAZVINI M.R.; HOSSEINI S.M.

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Journal: MODERN GENETICS JOURNAL (MGJ) Year:2015 | Volume:9 | Issue:4 (39) Page(s): 493-500

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Information Journal Paper

Title

EXPRESSION OF RECOMBINANT PROTEIN IN TOBACCO SEEDS

Author(s)

BAGHERI KH. | GHAZVINI M.R. | HOSSEINI S.M. | Issue Writer Certificate

Keywords

GUS GENEQ2

INDUCER SEQUENCEQ2

PROMOTERQ2

RECOMBINANT PROTEINQ2

TOBACCOQ2

Abstract

The low level expression of transgenes and their silence through successive generations are the most important factors limiting the production of RECOMBINANT PROTEINs in plants. Choosing an appropriate plant tissue for expression, and improving of the gene cassette could increase the expression of the transgene transferred to the plant. To address this problem, in this study, the increase of expression level of GUS GENE in TOBACCO seed was considered by designing and preparation of seed specific construct and transferring it into the TOBACCO explents. The signal sequence in upstream and KDEL in downstream of the GUS GENE were integrated as signals for protein retention in endoplasmic reticulum. The matrix attachment region (MAR) sequence was placed downstream of the GUS GENE to prevent silencing. The whole fragment was expressed under the control of Napin, the seed specific PROMOTER. This construct was transferred to A.tumefaciens LBA4404 and then used to transform TOBACCO leaf explants. Analysis of regenerated plants by using PCR and specific primers of nptII and GUS indicated the successful transfer of these genes to plantlets. The RT-PCR reaction results showed that nptII is transcribed in both tissues while GUS is only transcribed in the seed tissue. This finding is expected when considering that Nos PROMOTER (which controls the nptII transcription) is a constitutive and Napin is a seed specific PROMOTER. Expression of GUS in seeds of selected plants was investigated using SDS-PAGE and histochemical assay. The nearly 60 kDa band in the transgenic plant sample and also the results of histochemical assay indicate the expression of GUS GENE in transgenic seeds of TOBACCO.

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APA: Copy

BAGHERI, KH., GHAZVINI, M.R., & HOSSEINI, S.M.. (2015). EXPRESSION OF RECOMBINANT PROTEIN IN TOBACCO SEEDS. MODERN GENETICS JOURNAL (MGJ), 9(4 (39)), 493-500. SID. https://sid.ir/paper/116698/en

Vancouver: Copy

BAGHERI KH., GHAZVINI M.R., HOSSEINI S.M.. EXPRESSION OF RECOMBINANT PROTEIN IN TOBACCO SEEDS. MODERN GENETICS JOURNAL (MGJ)[Internet]. 2015;9(4 (39)):493-500. Available from: https://sid.ir/paper/116698/en

IEEE: Copy

KH. BAGHERI, M.R. GHAZVINI, and S.M. HOSSEINI, “EXPRESSION OF RECOMBINANT PROTEIN IN TOBACCO SEEDS,” MODERN GENETICS JOURNAL (MGJ), vol. 9, no. 4 (39), pp. 493–500, 2015, [Online]. Available: https://sid.ir/paper/116698/en

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