Cell and Tissue Journal

Abstract Aim: Papaver somniferum remains the sole commercial source for several pharmaceutical benzophenanthridine alkaloids from benzylisoquinoline branch alkaloids, which includes the narcotic analgesics codeine and the semi-synthetic drugs such as; oxycodone, buprenorphine and naltrexone. Although, most of the biosynthetic pathways genes of this alkaloid have been identified, the post-transcriptional regulation of these alkaloids pathway has not been completely determined. Virus induced gene silencing (VIGS) is a method for fast functional genomics, and in this study, this technique was used to investigate the silencing one of the most important genes in this alkaloid pathway.
Materials and Methods: In the current research VIGS technique was used for systematic reduction in the level of genes expression involves in benzylisoquinoline alkaloids pathway. For silencing of codeinone reductase gene, pTRV vector was used for cloning. The specific silencing of COR gene was evaluated in 2-3 weeks old leaves of Papaver somniferum using agro-infiltration method.
Results: Result of cloning was confirmed by using of different molecular methods such as enzyme digestion and also PCR. PCR technique was used for confirmation of transgenic plants in some transformed plants. Using of semi-quantitative PCR and real-time PCR showed that the level of reduction in transcription of COR gene was about 89 percent.
Conclusion: The obtained results confirmed that by application of RNA interference method, the level of COR gene expression was significantly reduced compared with control.

Abstract Aim: Our aim in this study was to analyze and quantify mRNA expression of SALL4 in mesencephalon, during different stages of chicken embryogenesis. Material and methods: In this experimental study, incubated Ross fertilized eggs were applied in 37°C-37.5°C in 60-65% humidified atmosphere after beginning of embryogenesis. Mesencephalon part of the brain tissue was collected from the eggs, daily. Total RNA extraction and cDNA synthesis was performed from resected tissues. The synthesized cDNA was used as template for quantitatively analysis of SALL4 mRNA expression by real-time PCR. Results: The Real-time PCR analysis of SALL4 mRNA expression in mesencephalon tissues indicated that the level of gene expression is significantly variable during embryogenesis. While the basal level of SALL4 mRNA expression was detected during the rest of embryogenesis, the maximum copy number of SALL4 mRNA was quantified on 19th day of chicken development. Conclusion: Having analyzed the level of SALL4 mRNA expression in different stages of chicken embryogenesis, we can extrapolate that a probable relationship may be existed between expression of SALL4 in nerve centers of mesencephalon brain and development of optic organs. Keywords: