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微生物催化D-葡萄糖与酪醇葡糖基转移合成红景天甙的初步研究 被引量:15
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作者 王梦亮 张芳 刘滇生 《催化学报》 SCIE CAS CSCD 北大核心 2006年第3期233-236,共4页
为了探讨微生物催化合成红景天甙的可能性,从红景天根系土壤中筛选出了五个菌株,比较了它们合成红景天甙的能力,确定F1菌株为合成红景天甙的出发菌株,经鉴定该菌株为米曲霉(Aspergillus oryzae).以D-葡萄糖和酪醇为底物,研究了底物浓度... 为了探讨微生物催化合成红景天甙的可能性,从红景天根系土壤中筛选出了五个菌株,比较了它们合成红景天甙的能力,确定F1菌株为合成红景天甙的出发菌株,经鉴定该菌株为米曲霉(Aspergillus oryzae).以D-葡萄糖和酪醇为底物,研究了底物浓度、反应时间、细胞浓度和pH对红景天甙产量的影响.结果表明,最适底物浓度为酪醇5 g/L,D-葡萄糖与酪醇的摩尔比为1∶1,酪醇浓度高于5 g/L不利于菌体生长.最佳反应时间为48 h,最适细胞浓度为150 g/L,最适pH为7.红景天甙最高产量为0.7 g/L. 展开更多
关键词 红景天甙 微生物 米曲霉 生物催化 酪醇 葡萄糖 葡糖基转移
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Functional analysis of a cotton glucuronosyltransferase promoter in transgenic tobaccos 被引量:4
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作者 Ai-Min Wu Shi-You Lv Jin-Yuan Liu 《Cell Research》 SCIE CAS CSCD 2007年第2期174-183,共10页
The 5' fragment (1 647 bp) of the cotton glucuronosyltransferase gene (GhGlcAT1) was transcriptionally fused to the β-glucuronidase (GUS) gene, and functionally analyzed for important regulatory regions contro... The 5' fragment (1 647 bp) of the cotton glucuronosyltransferase gene (GhGlcAT1) was transcriptionally fused to the β-glucuronidase (GUS) gene, and functionally analyzed for important regulatory regions controlling gene expression in transgenic tobacco plants. GUS activity analysis revealed that the full-length promoter drives efficient expression of the GUS gene in the root cap, seed coat, pollen grains and trichomes. Exposure of the transgenic tobacco to various abiotic stresses showed that the promoter was mainly responsive to the sugars (glucose and sucrose) as well as gibberellic acid. Progressive upstream deletion analyses of the promoter showed that the region from -281 to +30 bp is sufficient to drive strong GUS expression in the trichomes of shoot, suggesting that the 311 bp region contains all cis-elements needed for trichome-specific expression. Furthermore, deletion analysis also revealed that the essential cis-element(s) for sucrose induction might be located between -635 and -281 bp. In addition, sequence analysis of the regulatory region indicated several conserved motifs among which some were shared with previously reported seed-specific elements and sugarresponsive elements, while others were related with trichome expression. These findings indicate that a 1 647-bp fragment of the cotton GhGIcAT1 promoter contains specific transcription regulatory elements, and provide clues about the roles of GhGIcAT 1 in cotton fiber development. Further analyses of these elements will help to elucidate the molecular mechanisms regulating the expression of the GhGlcAT1 gene during fiber elongation. 展开更多
关键词 GLUCURONOSYLTRANSFERASE COTTON sugar-induced expression regulatory elements promoter activity transgenic tobacco
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Identification of Senescence-associated Protein DpXTH1 and its Gene Cloning in Dahlia Petals
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作者 张萍萍 阚雪芹 +3 位作者 李坚 陈驰 罗成洋 谈建中 《Agricultural Science & Technology》 CAS 2015年第7期1490-1493,共4页
[Objective] This study aimed to explore the molecular mechanism of senescence in ethylene-insensitive flowers. [Method] The dahlia petals were used as matedal, and the senescence-associated proteins were isolated and ... [Objective] This study aimed to explore the molecular mechanism of senescence in ethylene-insensitive flowers. [Method] The dahlia petals were used as matedal, and the senescence-associated proteins were isolated and identified using two-dimensional electrophoresis, mass spectrometry and an encoding gene was cloned using molecular biology techniques. [Result] In the two-dimensional elec- trophorogram of proteins from dahlia petals at building color, full flowering and flow- er senescence pedods, a total of 44 protein spots with differences in expression level more than two times were detected. From the 44 protein spots, xyloglucan (XTHs), a senescence-associated protein, was iso- lated and identified and its expression level was increased continuously with the senescence process of dahlia petals. By using the total RNA of dahlia petals as matedal and a pair of degenerate pdmers, the cDNA sequence of XTH gene was cloned by RT-PCR. The encoding region of XTH gene has a full length of 882 bp, encoding 293 amino acid residues, and is named as DpXTH1 (Accession number: HM053613.1). The cluster analysis showed that the amino acid sequence of DpXTH1 has high homology with those of XTHs in other plants. [Conclusion] The isolated and identified DpXTH1 from dahlia petals belonged to the XTH family in plants, and its biological function was associated with the senescence process and regulation of dahlia petals. 展开更多
关键词 DAHLIA PETALS Xyloglucan endotransglycosylase/hydrolase Senescence-associated protein Gene cloning
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Heterologous expression of active human undine diphosphate glucuronosyltransferase 1A3 in Chinese hamster lung cells 被引量:3
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作者 Ya-KunChen XinLi Shu-QingChen SuZeng 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第1期118-121,共4页
AIM: To obtain the active human recombinant uridine diphosphate glucuronosyltransferase 1A3 (UGT1A3) enzyme from Chinese hamster lung (CHL) cells.METHODS: The full-length UGT1A3 gene was amplified by reverse transcrip... AIM: To obtain the active human recombinant uridine diphosphate glucuronosyltransferase 1A3 (UGT1A3) enzyme from Chinese hamster lung (CHL) cells.METHODS: The full-length UGT1A3 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR)using total RNA from human liver as template. The correct fragment confirmed by sequencing was subcloned into the mammalian expression vector pcDNA3.1 (+), and the recombinant vector was transfected into CHL cells using a calcium phosphate method. Expressed UGT1A3 protein was prepared from CHL cells resistant to neomycin (G418). Then the protein was added into a reaction mixture for glucuronidation of quercetin. The glucuronidation activity of UGT1A3 was determined by reverse phase-high performance liquid chromatography (RP-HPLC) coupled with a diode array detector (DAD). The quercetin glucuronide was confirmed by hydrolysis with β-glucuronidase. Control experiments were performed in parallel. The transcriptions of recombinants were also determined by RT-PCR.RESULTS: The gene was confirmed to be an allele (UGT1A3-3) of UGT1A3 by DNA sequencing. The fragment was introduced into pcDNA3.1 (+) successfully. Several colonies were obtained under the selection pressure of G418.The result of RT-PCR showed transcription of recombinants in mRNA level. Glucuronidation assay and HPLC analysis indicated UGT1A3 expressed heterologously in CHL cells was in an active form, and one of the gulcuronides corresponding to quercetin was also detected.CONCLUSION: Correct sequence of UGT1A3 gene can be obtained, and active UGT1A3 enzyme is expressed heterologously in CHL cells. 展开更多
关键词 Uridine Diphosphate Glucuronosyltransferase 1A3 LUNG
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No association between cyclooxygenase-2 and uridine diphosphate glucuronosyltransferase 1A6 genetic polymorphisms and colon cancer risk 被引量:11
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作者 Cheryl L Thompson Sarah J Plummer +4 位作者 Alona Merkulova Iona Cheng Thomas C Tucker Graham Casey Li Li 《World Journal of Gastroenterology》 SCIE CAS CSCD 2009年第18期2240-2244,共5页
AIM:To investigate the association of variations in the cyclooxygenase-2 (COX2) and uridine diphosphate glucuronosyltransferase 1A6 (UGTIA6) genes and non-steroidal anti-inflammatory drugs (NSAIDs) use with ris... AIM:To investigate the association of variations in the cyclooxygenase-2 (COX2) and uridine diphosphate glucuronosyltransferase 1A6 (UGTIA6) genes and non-steroidal anti-inflammatory drugs (NSAIDs) use with risk of colon cancer.METHODS: NSAIDs, which are known to reduce the risk of colon cancer, act directly on COX2 and reduce its activity. Epidemiological studies have associated variations in the COX2 gene with colon cancer risk, but others were unable to replicate this finding. Similarly,enzymes in the UGT1A6 gene have been demonstrated to modify the therapeutic effect of NSAIDs on colon adenomas. Polymorphisms in the UGTIA6 gene have been statistically shown to interact with NSAID intake to influence risk of developing colon adenomas, but not colon cancer. Here we examined the association of tagging single nucleotide polymorphisms (SNPs) in the COX2 and UGTIA6 genes, and their interaction with NSAID consumption, on risk of colon cancer in a population of 422 colon cancer cases and 481 population controls.RESULTS: No SNP in either gene was individually statistically significantly associated with colon cancer, nor did they statistically significantly change the protective effect of NSAID consumption in our sample. Like others, we were unable to replicate the association of variants in the COX2 gene with colon cancer risk (P 〉 0.05),and we did not observe that these variants modify the protective effect of NSAIDs (P 〉 0.05). We were able to confirm the lack of association of variants in UGT1A6 with colon cancer risk, although further studies will have to be conducted to confirm the association of these variants with colon adenomas.CONCLUSION: Our study does not support a role of COX2 and UGTIA6 genetic variations in the development of colon cancer. 展开更多
关键词 Uridine diphosphate glucuronosyltransferase 1A6 CYCLOOXYGENASE-2 Non-steroidal anti-inflammatorydrugs Colon cancer Genetic association studies Singlenucleotide polymorphisms
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Two unrelated patients with rare Crigler-Najjar syndrome type I:two novel mutations and a patient with loss of heterozygosity of UGT1A1 gene 被引量:2
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作者 Yan LI Yu-jin QU +8 位作者 Xue-mei ZHONG Yan-yan CAO Li-min JIN Jin-li BAI Xin MA Yu-wei JIN Hong WANG Yan-ling ZHANG Fang SONG 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2014年第5期474-481,共8页
Cdgler-Najjar syndrome type Ⅰ (CN-I) is the most severe type of hereditary unconjugated hyperbilirubinemia. It is caused by homozygous or compound heterozygous mutations of the UDP-glycuronosyltransferase gene (UG... Cdgler-Najjar syndrome type Ⅰ (CN-I) is the most severe type of hereditary unconjugated hyperbilirubinemia. It is caused by homozygous or compound heterozygous mutations of the UDP-glycuronosyltransferase gene (UGT1A1) on chromosome 2q37. Two patients clinically diagnosed with CN-I were examined in this paper. We sequenced five exons and their flanking sequences, specifically the promoter region of UGT1A 1, of the two patients and their parents. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine the UGT1A1 gene copy number of one patient. In patient A, two mutations, c.239_245delCTGTGCC (p.Pro80HisfsX6; had not been reported previously) and c.1156G〉T (p.Va1386Phe), were identified. In patient B, we found that this patient had lost heterozygosity of the UGTIA1 gene by inheriting a deletion of one allele, and had a novel mutation c.1253delT (p.Met418ArgfsX5) in the other allele. In summary, we detected three UGTIA 1 mutations in two CN-I patients: c.239_ 245delCTGTGCC (p.Pro80HisfsX6), c.1253delT (p.MeH18ArgfsX5), and c.1156G〉T (p.Va1386Phe). The former two mutations are pathogenic; however, the pathogenic mechanism of c.1156G〉T (p.Va1386Phe) is unknown. 展开更多
关键词 Crigler-Najjar syndrome type (CN-I) HYPERBILIRUBINEMIA UDP-glycuronosyltransferase gene (UGT1A 1) Mutation Loss of heterozygosity
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