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Fine mapping and candidate gene prediction of the photoperiod and thermo-sensitive genic male sterile gene pms1(t) in rice

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2011年第6期12卷 436-447页

作者： Yuan-fei ZHOU Xian-yin zhang Qing-zhong xue Department of Agronomy College of Agriculture and Biotechnology Zhejiang University Hangzhou 310029 China

Pei＇ai64S, an indica sterile variety with photoperiod and thermo-sensitive genic male sterile （PTGMS） genes, has been widely exploited for commercial seed production for ＂two-line＂ hybrid rice in China. One PTGMS gene from Pei＇ai64S, pmsl（t）, was mapped by a strategy of bulked-extreme and recessive-class approach with simple sequence repeat （SSR） and insert and deletion （In-Del） markers. Using linkage analysis for the F2 mapping population consisting of 320 completely male sterile individuals derived from a cross between Pei＇ai64S and 93-11 （indica restorer） lines, the pmsl（t） gene was delimited to the region between the RM21242 （0.2 cM） and YF11 （0.2 cM） markers on the short arm of chromosome 7. The interval containing the pmsl（t） locus, which was co-segregated with RM6776, is a 101.1 kb region based on the Nipponbare rice genome. Fourteen predicted loci were found in this region by the Institute for Genomic Research （TIGR） Genomic Annotation. Based on the function of the locus LOC_Os07g12130 by bioinformatics analysis, it is predicted to encode a protein containing a Myb-like DNA-binding domain, and may process the transcript with thermosensory response. The reverse transcription-polymerase chain reaction （RT-PCR） results revealed that the mRNA levels of LOC_Os07g12130 were altered in different photoperiod and temperature treatments. Thus, the LOC_Os07g12130 locus is the most likely candidate gene for pmsl（t）. These results may facilitate not only using the molecular marker assisted selection of PTGMS genes, but also cloning of the pmsl（t） gene itself.

关键词： Super hybrid rice pmsl(t) Photoperiod and thermo-sensitive male sterile gene Genetic mapping

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Preparation and Anti-oxidative Effects of Corn Peptides

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Chemical Research in Chinese Universities 2002年第3期18卷 299-302页

作者： XU Li zhang xue-zhong GUO Yi REN Qiao WU Xiao-xia Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education Jilin UniversityChangchun 130021P.R.China

Corn peptides(CP) were prepared from zein via proteolysis by means of a type of alkaline protease, Alcalase, at 50 ℃ and pH 8.0. The anti oxidative activity of the hydrolysate was examined. It was found that the anti oxidative activity reached the maximum at the fourth hour of the hydrolysis and then decreased. The effect of the enzyme dosage on the solubility of corn protein, the dependence of hydrolysis time on the degree of the hydrolysis(DH) and molecular weight distribution were examined respectively. 30% DH was controlled on the basis of anti oxidative activity. The hydrolysate was fractionated on a Sephadex G 15 column. The anti oxidative activities were examined for all the fractions. The main fraction for anti oxidative activity was collected and characterized by the methods of amino acid analysis, the measurement of average chain length and etc .. The main M W distribution of the main fraction was 300-600 Daltons. The anti oxidative properties of the small peptides in vitro were studied. It was found that they could inhibit the auto oxidation of pyrogallol and the oxidation of hemoglobin. The hemolysis degree of erythrocyte caused by hydroxyl free radicals was decreased greatly, indicating a protective effect of the anti oxidative peptides on the cell membrane damage of erythrocyte.

关键词： Corn Peptides Preparation Anti oxidative effect

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Chemo-enzymatic Synthesis of Z-Asp-Val-Tyr-NH_2——A Precursor Tripeptide of Thymopentin

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Chemical Research in Chinese Universities 2010年第6期26卷 942-947页

作者： GUO Yi zhang Yong-xia JIANG Xin zhang xue-zhong WU Yi XU Li Key Laboratory for Molecular Enzymology and Engineering Ministry of Education Jilin University Changchun 130012 P. R. China College of Pharmacy Jilin University Changchun 130021 P. R. China

Z-Asp-Val-Tyr-NH2,a precursor tripeptide of thymopentin（TP-5）,was successfully synthesized by a combination of chemical and enzyme ***,the precursor dipeptide Val-Tyr-NH2 was synthesized by a novel chemical method in four steps including the preparation of NCA-Val,the esterification of L-tyrosine and the ammonolysis of L-tyrosine ethyl ester,and the formation of Val-Tyr-NH2 from NCA-Val and ***,a thermolysin,thermoase PC 10F was used to catalyze the formation of Z-Asp-Val-Tyr-NH2 in an aqueous/organic solvent diphase system with Z-Asp-OH and Val-Ty-NH2 as the substrates under thermodynamic *** optimum conditions were pH=6.5,40 ℃,thermoase 100 mg,in 2-（N-morpholino） ethanesulfonic acid MES/NaOH（containing 5 mmol/L CaCl2） of n-butanol system（15/85,volume ratio） for 20 h in a maximum yield of 27.02%.

关键词： Thymopentin Peptide synthesis Thermoase Organic solvent

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A new protoberberine alkaloid from Corydalis yanhusuo ***

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Chinese Chemical Letters 2009年第8期20卷 955-957页

作者： Tian Tian Hu xue zhang Shi zhong Ma Xin Sheng Yao School of Traditional Chinese Materia Medica Shenyang Pharmaceutical University Shenyang 110016 China

A new protoberberine alkaloid, named 5,6-dihydro- 10-hydroxy-2,3,9-trimethoxy- 13-methyldibenzo[α,g]quinoliziniurn （1） was isolated from the 60% ethanol extract of the tubers of Corydalis yanhusuo W. T. Wang, together with a new natural product, 13methylpalmatrubine （2）. Their structures were established by spectroscopic methods. 2009 Xin Sheng Yao. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All fights reserved.

关键词： Corydalis yanhusuo W. T. Wang Protoberberine alkaloid 13-Methylpalmatrubine

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Studies on the Conformation of Poly (1-naphthol) Assembled by Horseradish Peroxidase in Aqueous Micelle

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Chemical Research in Chinese Universities 1998年第2期14卷 59-62页

作者： CHEN Song-ming zhang xue-zhong CHAI Xiang-dong LI Tie-jin Department of Chemistry the State Key Lab of Enzyme EngineeringJilin UniversityChangchun130023

FTIR, UV-visible , fluorescence spectra and calculation of CPK model indicated that poly(1-naphthol) synthesized and assembled by horseradish peroxidase(HRP) in aqueous micelle was constructed with orderly helical conformation, which was the lowest energy conformation of the polymer under this condition and resulted in an advanced c-conjugated system.

关键词： Conformation Poly(1naphthol) Horseradish peroxidase Aqueous micelle

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Protease-catalyzed Synthesis of Bz-Arg-Gly-Asp-OMe in Full Aqueous Medium

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Chemical Research in Chinese Universities 2007年第6期23卷 693-696页

作者： HOU Rui-zhen LI Hong-mei LIU Yun-jia zhang Long XU Li zhang xue-zhong Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education Jilin University Changchun 130021 P. R. China College of Chemical Engineering Changchun University of Technology Changchun 130012 P. R. China College of Food Engineering Jilin Agricultural University Changchun 130015 P. R. China

Synthesis of N-benzoyl-argininylglycylasparagine methyl ester（ Bz-Arg-Gly-Asp-OMe）, a precursor tripeptide of Arg-Gly-Asp） was catalyzed by papain under kinetic control, at alkaline pH, in a full aqueous medium. The substrates were N-benzoyl-argininylglycine ethyl ester and asparagine dimethyl ester. An aqueous solution of 0. 1 mol/L KCl/NaOH containing 8 mmol/L EDTA and 2 mmol/L DTT was selected as the reaction medium. The synthesized hydrophilic tripeptide was soluble in the reaction medium during the reaction process, however, the secondary hydro- lysis of the tripeptide product was not considerable. The effects of different factors, including water content, temperature, reaction time, and molar ratio of the substrates, on the yield of Bz-Arg-Gly-Asp-OMe were examined. The optimal reaction conditions were 0. 05 mol/L Bz-Arg-Gly-OEt and 0. 15 mol/L Asp（ -OMe）2·HCl in 0. 1 mol/L KCl/ NaOH solution（pH 8.5）, at 40 ℃, and a reaction time of 60 min, with a maximum conversion yield of 62.4%.

关键词： RGD Peptide synthesis Papain Kinetic control

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A Study on Anti-oxidative Activity of Soybean Peptideswith Linoleic Acid Peroxidation Systems

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Chemical Research in Chinese Universities 2006年第2期22卷 205-208页

作者： XU Li LI Hong-mei HUANG Yi-bing WANG Hua NIE Guang-jun zhang xue-zhong Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education Jilin University Changchun 130021 P. R. China College of Food Engineering Jilin Agricultural University Changchun 130018 P. R. China China-Japan Union Hospital of Jilin University Changchun 130033 P. R. China

Soybean bioactive peptides（SBPs） were prepared from the isolated soybean protein by proteolysis with an alkaline protease, alcalase, at 50℃ and pH = 8.0. The dependence of hydrolysis time on hydrolysis degree and molecular weight distribution were examined. The hydrolysate was fractionated on a Sephadex G-25 column and the anti-oxidative activities of the fractions were detected by the method of pyrogallol auto-oxidation. The average chain length of soybean peptides that have anti-oxidative activity was estimated to be about 7. The anti-oxidative properties of the soy- bean peptide were also studied by using linoleic acid peroxidation systems. The optimal condition of the peroxidation system was set up, Vc/Cu^2 ＋ as the inducer at pH = 7.4 and 25 ℃. In addition, soybean peptides show higher antioxidative activity compared with GSH.

关键词： Soybean bioactive peptides Anti-oxidative activity Linoleic acid peroxidation system

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RGDC Peptide Modified Quantum Dots Labelling and Imaging of Tumor Cells

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Chemical Research in Chinese Universities 2011年第5期27卷 832-835页

作者： GUO Yi LI Chun-rong SHEN Huai-bin zhang xue-zhong LI Lin-song YU Qian XU Li Key Laboratory for Molecular Enzymology and Engineering Ministry of Education College of Life Science Jilin University Changchun 130012 P. R. China Department of Drug Administration China-Japan Union Hospital of Jilin University Changchun 130033 P. R. China Key Laboratory for Special Functional Materials Ministry of Education Henan University Kaifeng 475004 P. R. China

The labelling and imaging of tumor cells were investigated via arginine-glycine-aspartic acidcysteine（RGDC） peptide-labelled quantum dots（QDs）. The results show that RGDC modified QDs can label SMMC-7721 tumor cells and adhere to cellular membrane. In constrast, the unmodified QDs are mainly dispersed around the cell. We also found that the RGDC-QDs can penetrate into the cell at 2 h of incubation. After 6 h of incubation, RGDC-QDs can accumulate in a unique intracellular region.

关键词： Arginine-glycine-aspartic acid-cysteine(RGDC) peptide Quantum dot Fluorescence labelling

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Chemoenzymatic Synthesis of Cellular Adhesion Tripeptide RGD Precursor in Organic Media

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Chemical Research in Chinese Universities 2006年第6期22卷 721-726页

作者： WANG Hua ZHAO Mi-feng PU Li-mei DING Wei HUANG Yi-bing HOU Rui-zhen ZENG Hong-bin XU Li zhang xue-zhong Key Laboratory of Molecular Enzymology and Engineering Jilin University Changchun 130021 P. R. China China-Japan Union Hospital Jilin University Changchun 130033 P. R. China First Hospital Jilin University Changchun 130021 P. R. China

Chemoenzymatic synthesis of tripeptide Bz-RGD-（OEt）2 was conducted in this study. First, the free dipeptide Gly-Asp was synthesized via a novel chemical method, wherein only L-aspaxtic acid was used and was followed by the esterification of Gly-Asp. The formation of the linkage between the third amino acid Bz-Arg-OEt and Gly-Asp-（OEt）2 was completed by using the enzymatic method in organic media. The effects of several factors such as pH, the water content, triethylamine （TEA）, the molar ratio of the substrates, and the reaction time on the yield of Bz-RGD- （OEt）2 were examined. It was obtained that the optimum conditions for Bz-RGD-（OEt）2 synthesis in an ethanol/ Tris-HC1 buffer system （ volume ratio 93： 7 ） were as follows ： pH = 8.0 ; temperature, 30 ℃ ; reaction time, 7 h. The tripeptide yield was 75.2%.

关键词： RGD Synthesis Trypsin Organic solvent

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Inhibition of Growth of Human Ileocecal Adenocarcinoma Cells HCT-8 and Inducing Apoptosis by Different RGD-containing Peptides

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Chemical Research in Chinese Universities 2008年第4期24卷 481-486页

作者： WANG Hua YANG Shao-juan GAO Shuo-hui HUANG Yi-bing LI Jing CAI Ming-jun XU Li zhang xue-zhong Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education Jilin University Changchun 130021 P. R. China China-Japan Union Hospital of Jilin University Changchun 130033 P. R. China

Human ileocecal adenocarcinoma cells HCT-8 were treated with RGD-containing cellular adhesion peptides including RGD, RGD（NH2）2（i.e., RGE-NH2）, RGDS, and RGDS-NH2. MTT assay was prepared to examine their inhibiting effects on HCT-8 cells after treatment. The methods including Haematoxylin and Eosin（HE） staining, transmission electron microscopy（TEM）, immunohistochemistry, flow cytometry, and Reverse TranscriptionPolymerase Chain Reaction（RT-PCR） were used to observe the morphology of the apoptotic cells and analyze the mechanism of apoptosis. The experimental results indicate that RGD-containing cellular adhesion peptides can inhibit the growth and proliferation of tumor HCT-8 cells in a dose-dependent manner and induce the apoptosis of HCT-8 cells. At the same time, the high conservative property of RGD was confirmed again.

关键词： RGD HCT-8 cells Cellular adhesion Cell apoptosis

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