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Over-expression of StLCYb increases b-carotene accumulation in potato tubers
Xiao-yan Song,Wen-jiao Zhu,Rui-min Tang,Jing-hui Cai,Min Chen,Qing Yang 한국식물생명공학회 2016 Plant biotechnology reports Vol.10 No.2
Lycopene b-cyclase (LCYb) is involved in the first step of the b-branch synthetic pathway of carotenoids from lycopene in plants. In this study, to explore the possibility of regulating b-carotene synthesis via the b-branchspecific pathway in potato, StLCYb gene was first isolated from potato cultivar Desiree, and its open reading frame was 1503 bp long without intron. The protein sequence of StLCYb showed high similarity with that of LCYbs in other species such as SlLCYb1, CaLCYb, NtLCYb and ApLCYb. StLCYb was expressed in all tissues and the highest level was observed in tubers followed in flowers, and the lowest level was in roots. HPLC detected an about 1.5–1.9 times increase in b-carotene content of transgenic potato tubers, in which the gene StLCYb was overexpressed, compared with the wild-type control. Besides, the expression levels of carotenoid-related genes StPSY, StPDS, StZDS, StCHYb and StZEP transcripts in the transgenic lines were significantly higher than in the wild-type control, which implied a positive regulation in promoting carotenoid synthesis. All these results suggest that b-carotene content in potato tubers could be regulated by modulating StLCYb expression.
Wu, Guo-Qiu,Liu, Nan-Nan,Xue, Xiu-Lei,Cai, Li-Ting,Zhang, Chen,Qu, Qing-Rong,Yan, Xue-Jiao Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.10
Background: This study was aimed to establish a novel method to simultaneously detect expression of four genes, ribonucleotide reductase subunit M1(RRM1), X-ray repair cross-complementing gene 1 (XRCC1), thymidylate synthase (TS) and class III ${\beta}$-tubulin (TUBB3), and to assess their application in the clinic for prediction of response of non-small cell lung cancer (NSCLC) to chemoradiotherapy. Materials and Methods: We have designed four gene molecular beacon (MB) probes for multiplex quantitative real-time polymerase chain reactions to examine RRM1, XRCC1, TUBB3 and TS mRNA expression in paraffin-embedded specimens from 50 patients with advanced or metastatic carcinomas. Twenty one NSCLC patients receiving cisplatin-based first-line treatment were analyzed. Results: These molecular beacon probes could specially bind to their target genes in homogeneous solutions. Patients with low RRM1 and XRCC1 mRNA levels were found to have apparently higher response rates to chemoradiotherapy compared with those with high levels of RRM1 and XRCC1 expression (p<0.05). The TS gene expression level was not significantly associated with chemotherapy response (p>0.05). Conclusions: A method of simultaneously detecting four molecular markers was successfully established and applied for evaluation of chemoradiotherapy response. It may be a useful tool in personalized cancer therapy.
Hong-juan Zhang,Wei-guo Zhang,Zhi-yuan Wang,Yue-ping Zhan,Li-sheng Xu,Jun-zhong Liu,Qian Liu,Qing-cai Jiao 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.2
In order to investigate the catalytic mechanism of Escherichia coli γ-glutamyltranspeptidase, ten para- and meta- substituted γ-glutamyl anilides were chemically prepared and employed as substrates to synthesize L-theanine to assay the activity of γ-glutamyltranspeptidase. The reaction was optimized for γ-glutamyl-p-nitroanilide. Key factors such as substrate specificity, pH, temperature, and the substrate mole ratio were all investigated. Kinetic studies of the acyl transfer reaction were described and the Hammett plot was constructed. This study indicated that the ratelimiting acylation reaction of γ-glutamyltranspeptidase can apparently be accelerated by either the electron-withdrawing or electron-donating substituents of γ-glutamyl anilides. The reaction could be catalyzed by the general acid and carboxy of Asp-433 or phenolic hydroxyl Tyr-444 may be the acid by autodock simulation for all prepared γ-glutamyl anilides.