1.Construction of RNAi Expression Vector by Fusion Gene Fragments of BBE and COR from Papaver nudicarule
Qianqian LIANG ; Jinwen ZHANG ; Yujie WEI ; Wangtian WANG ; Qingxiang HE ; Yaohu LEI
China Biotechnology 2006;0(11):-
The codeinone reductase gene (cor) and the berberine bridge enzyme gene (bbe) were cloned from young leaf of opium poppy(Papaver somniferum L) by RT-PCR and the coding sequences of these gene were analyzed. The result demonstrated that the cloned COR gene sequence was highly homologous to the other COR gene family members showing 98.96% identity to COR1.1. The cloned BBE gene sequence was 94.84% identified with the released BBE genes in GenBank previously. Based on the cDNA sequences of COR and BBE,two fragments about 400~500 bp from each gene with lower identity among them were cloned. The fusion gene BC(744 bp) is fused by the PCR technique. Then the promoter CaMV 35S driven,containing'forward BC fusion fragments-reverse pdk intron-reverse BC fusion fragments',plant siRNA expression vector were constructed based on the vectors pHANNIBAL and pART27. Inhibition efficiency of the expression vector to the morphine synthesis was studied by transforming papaver nudicarule.The work will lay the foundation for breeding a low morphine and high thebaine poppy.
2.Hydrolytic property and solution stability of NAMI derivative containing nicotinamide.
Yaohu LIANG ; Wei BI ; Han YAN ; Guogang LIANG
China Journal of Chinese Materia Medica 2012;37(6):818-823
OBJECTIVETo study the influence of ligand structure on hydrolysis and solution stability of NAMI derivatives.
METHODNAMI type compound 1, trans- [RuCl4 (DMSO) (nica)] Na x 2DMSO (nica, nicotinamide) were prepared. Their hydrolytic mechanism, kinetics and stability were investigated by UV-Vis spectrophotometer.
RESULTSimilar to NAMI, compound 1 undergoes two well-separated steps chloro-hydrolysis (I chloro-hydrolysis and II chloro-hydrolysis) (step reaction) in pH 7.4 buffer solution; while dimethyl sulfoxide (DMSO) hydrolyze in pH 5.00 acetic buffer solution. The k(obs) and t1/2 for each hydrolytic reaction were determined.
CONCLUSIONThe stability of compound 1 in acidic solution is much more stable than that of in neutral solution. Nicotinamide in place of imidazole can decrease chloro hydrolytic rate of NAMI derivatives obviously, while the influence on the DMSO hydrolytic process is not so remarkable.
Dimethyl Sulfoxide ; chemistry ; Drug Stability ; Hydrolysis ; Imidazoles ; chemistry ; Kinetics ; Ligands ; Niacinamide ; chemistry ; Ruthenium ; chemistry ; Solutions ; chemistry