Isolation, cloning and transient silencing of BBE1 gene using virus induced gene silencing technique in Iranian genotypes of Papaver somniferum L.

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SOHRABI S.M.; ISMAILI A.; NAZARIAN FIROUZ ABADI F.

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Journal: Journal of Molecular and Cellular Researches Year:2018 | Volume:31 | Issue:3 Page(s): 364-375

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Information Journal Paper

Title

Isolation, cloning and transient silencing of BBE1 gene using virus induced gene silencing technique in Iranian genotypes of Papaver somniferum L.

Author(s)

SOHRABI S.M. | ISMAILI A. | NAZARIAN FIROUZ ABADI F. | Issue Writer Certificate

Keywords

poppyQ1

gene silencingQ2

pTRV plasmidQ1

real-time PCRQ1

Abstract

The poppy (Papaver somniferum L. ) is one of the oldest medicinal plant in the world. It remains the only commercial source for the important narcotic analgesics and semisynthetic derivatives such as oxycodone and naltrexone. Several other benzylisoquinoline alkaloids with potent pharmacological properties including papaverine, noscapine and sanguinarine also produce with this plant. BBE1 is one of key genes in biosynthesis of benzylisoquinoline alkaloids and catalyzes first step in production of noscapine and sanguinarine alkaloids. In the present study, the full length CDS of BBE1 gene was isolated from poppy plant using specific primers. Then, a specific fragment, corresponding to the BBE1 CDS was selected as silencing fragment and cloned into pTZ57R/T plasmid. In the next step, silencing fragment was cloned into pTRV plasmid and resulting constructs (pTRV2-BBE1 and pTRV2-Empty) along with helper plasmid (pTRV1) were transferred to Agrobacterium and infiltrated into young leaves. Result showed a 1608 bp DNA fragment for BBE1 gene. PCR analysis using CP specific primers showed presence of CP transcripts in the infiltrated plants. real-time PCR analysis showed transcript reduction (about 73%) in the transgenic plant (infiltrated with pTRV2-BBE1) compared with control plants (infiltrated with pTRV2-Empty and non-infiltrated).

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APA: Copy

SOHRABI, S.M., ISMAILI, A., & NAZARIAN FIROUZ ABADI, F.. (2018). Isolation, cloning and transient silencing of BBE1 gene using virus induced gene silencing technique in Iranian genotypes of Papaver somniferum L.. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), 31(3 ), 364-375. SID. https://sid.ir/paper/248556/en

Vancouver: Copy

SOHRABI S.M., ISMAILI A., NAZARIAN FIROUZ ABADI F.. Isolation, cloning and transient silencing of BBE1 gene using virus induced gene silencing technique in Iranian genotypes of Papaver somniferum L.. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY)[Internet]. 2018;31(3 ):364-375. Available from: https://sid.ir/paper/248556/en

IEEE: Copy

S.M. SOHRABI, A. ISMAILI, and F. NAZARIAN FIROUZ ABADI, “Isolation, cloning and transient silencing of BBE1 gene using virus induced gene silencing technique in Iranian genotypes of Papaver somniferum L.,” JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), vol. 31, no. 3 , pp. 364–375, 2018, [Online]. Available: https://sid.ir/paper/248556/en

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