a. m. latifi

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.
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.
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.
The obtained results confirmed that by application of RNA interference method, the level of COR gene expression was significantly reduced compared with control.

Lateral flow assay method is a test for rapid detection of morphine that h-as many applications in identification of ad-dicts. However، detection level of the sys-tem has limited its application. This rese-arch examined how to improve the detec-tion level and sensitivity of this system.

pH of conjugation، OD of conjugated material and concentr-ation of morphine-BSA in the test band w-ere effective on the detection level of this system. To determine optimum pH، a com-mercial kit (Gold in a box) was used. By applying various concentrations of morp-hine solution، and considering a constant concentration of morphine-BSA، the opt-imal OD was set. Then، the optimal conc-entration of morphine-BSA was obtained for a fixed amount of OD. Effect of block-ing the conjugated pad and addition of su-gar to the conjugated substance were also studied.

Optimum values of pH and OD was pH=8. 4 and OD=1. Optimum conce-ntration of morphine-BSA in the test band was obtained to be 0. 5 mg/ml. Cut-off value of optimized kit attained to 25 ng/ml. After comparing the results with a standard kit، sensitivity and specificity of the optimized kits were 100 % and 99%، respectively. Discussion &

Finding and opt-imizing the effective factors on the perf-ormance of diagnostic lateral flow method can be an effective strategy to increase the sensitivity and reduce the detection level of this system