title: Transient Expression of cor Gene in Papaver somniferum.
abstract: BACKGROUND
  Papaver somniferum is the commercial source of morphine and codeine. The isolation of effective genes involved in the morphine biosynthesis of P. somniferum is very important in the production of specific metabolites achieved using metabolic engi-neering techniques. In this pathway, the key enzyme COR is involved in the conversion of codeinone to codeine and morphinone to morphine.
  
  
  METHODS
  the gene encoding of this enzyme was isolated using primers designed on the base of gene sequence available on (NCBI) for P. somniferum. This gene correct size around (960 bp) was first subcloned into pTZ57RIT vector then cloned into expression vectors (pBI121) between BamHI and SacI sites to allow the expression of cor gene driven by the cauliflower mosaic virus 35S pro-moter. The result was confirmed through different molecular methods e.g. PCR and en-zyme digestion by BamHI and SacI. The recombinant plasmid was transformed into the E. coli strain DH5α using a freeze-thaw method. Having selected positive colones on selection medium, plasmid was extracted by miniprep method and recombinant plasmids were selected based on PCR and digestion. The construct was then mobilized in Agrobacterium tumefaciens C58/pGV3850 (KmR RifR). After gene transformation to P. somniferum plants, the agroinfiltration method was also used for transient expression of COR enzyme.
  
  
  RESULTS
  evaluation results showed that morphine and codeine were detectable in the leaves of transgenic plants containing cor transgene and there was significant difference in the final production. After completing this experiment for three times, results showed that in 11 sets from 15 sets of leaves experiment tested, main alkaloids (codeine, morphine, papaverin, noscapine and thebaine) were detectable.
  
  
  CONCLUSIONS
  Whereas no signal was detected in non-infiltrated control leaves or in leaves infiltrated with non-recombinant bacteria for morphine and codeine, others such as thebaine and papaverine were detectable.
authors.affiliation: Department of Horticulture, Faculty of Agriculture, Urmia University, Urmia, Iran.
authors.forename: Bahman
authors.initials: B
authors.lastname: Hosseini
authors.forename: Farajollah
authors.initials: F
authors.lastname: Shahriari-Ahmadi
authors.forename: Haleh
authors.initials: H
authors.lastname: Hashemi
authors.forename: Mohammad-Hasan
authors.initials: MH
authors.lastname: Marashi
authors.forename: Mahdi
authors.initials: M
authors.lastname: Mohseniazar
authors.forename: Alireza
authors.initials: A
authors.lastname: Farokhzad
authors.forename: Masoume
authors.initials: M
authors.lastname: Sabokbari
keywords: Agroinfiltration; Biosynthesis Pathway; COR Enzyme; Papaver somniferum