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Loss of Posterior Silk Gland Transcription Specificity of Fibroin Light Chain Promoter due to Absence of 41 bp Sequence Containing Possible Inhibitor Binding Sites

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Acta Biochimica et Biophysica Sinica 2005年第12期37卷 819-825页

作者： Ting-Qing GUO~(1,2) Jian-Yang WANG~(1,2) Sheng-Peng WANG~(1,2.3) Xiu-Yang GUO~(1,2) Ke-Wei HUANG3~3 Jun-Ting HUANG~3 Chang-De LU~11 institute of Biochemistry and Cell Biology,Shanghai institutes for Biological sciences.chinese academy of sciences.Shanghai 200031.china 2 Graduate School of the chinese academy of sciences.Shanghai 200031.china 3 Serictdtu research institute.chinese academy of agricultural sciences,zhenjiang 212.18.china institute of Biochemistry and Cell Biology Shanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200031China Graduate School of the chinese academy of sciences Shanghai 200031China sericultural research institute Chinese sericultural research institute Chinese Academy of Agricultural SciencesZhenjiang 212018China

The gene encoding fibroin light chain protein (FibL) is specifically expressed in the posteriorsilk gland of silkworm and repressed in other *** binding sites of several transcription factorsrevolved in the silk gland transcription specificity of fibl promoter have been recognized,including SGFB,PSGF and *** we report the leak expression of the enhanced green fluorescent protein (EGFP)reporter gene in tissues other than the posterior silk gland in vivo when under the control of a shortened fiblpromoter with deletion of the 5′terminal 41 bp sequence,which is located at -650 nt to -610 nt upstream *** transcription starting *** of silk gland specificity of the promoters was performed by observationof green fluorescence in tissues of silkworm larvae following inter-haemocoelic injection of recombinantAutographa californica multiple nuclear polyhedrosis virus carrying the EGFP reporter gene controlled bydifferent lengths of fibl *** results indicated that availability of the binding sites of several knownfactors,including SGFB,PSGF and BMFA,is not sufficient for intact silk gland transcription specificity offibl promoter,and there are possible inhibitor binding sites in the 41 bp sequence (-650 nt to -610 nt)upstream of the transcription starting site which may be required to repress the activity of fibl promoter inother tissues.

关键词： promoter specificity fibroin light chain silkworm Bombyx mori recombinant AcMNPV

Sequence analysis and expression of the calcium exchanger gene,MCAX-1,in mulberry（Morus alba L.）

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Sequence analysis and expression of the calcium exchanger ge...

现代分子植物育种与粮食安全研讨会

作者： Liu Zhaoyue1,2,Zhao Weiguo 1,2,Zhang Yinghua1,2,Tong Wei1,2,Fang Rongjun1,2,Li Long 1,2 * (1 College of biological and Environmental Engineering,jiangsu University of Science and technology,zhenjiang jiangsu 212.18,china 2 sericultural research institute,chinese academy of agricultural sciences,zhenjiang jiangsu 212.18,china)

For the sake of keeping the balance of plant cells,Calcium,as a necessary nutrition element to plant growth development,can get into the cytoplast by calcium channel and out of it by Ca2+-ATPase and calcium exchanger to hold a low calcium ***,calcium exchanger is related to resisting adversity of plants,to make the calcium mediated signal system unblocked and regulate adaptability accurately and ***,people are more and more focusing on the study of the calcium exchanger *** full-length cDNA that is cloned through RACE and RT-PCR methods,named as MCAX-1,cDNA fragments of which has been foundby measuring the function comments of ESTs sequence in mulberry,is 1784bp by the sequence *** the research focuses on gene expressions and prokaryotic expressions of the calcium exchanger gene under low temperature,drought,and salt stress conditions to find out gene expression patterns and responses of MCAX-1 in different *** will help us provide the theoretical basis and strong applications to excavate excellent gene resources and resistant molecular breeding in mulberry.

关键词： Mulberry Calcium exchanger gene(MCAX-1) Gene cloning Sequence analysis Gene expression

Molecular Cloning,Sequence Analysis,and Expression of the Methionine sulfoxide reductase Gene （MSR） in Mulberry

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Molecular Cloning,Sequence Analysis,and Expression of the Me...

现代分子植物育种与粮食安全研讨会

作者： Zhang Yinghua1,2,Liu Zhaoyue1,2,Tong Wei1,2,Fang Rongjun1,2,Liu Li1,2,Zhao Weiguo 1,2 * (1 College of biological and Environmental Engineering,jiangsu University of Science and technology,zhenjiang jiangsu 212.18,china 2 sericultural research institute,chinese academy of agricultural sciences,zhenjiang jiangsu 212.18,china )

A full-length cDNA sequence encoding methionine sulfoxide reductase （MSR） from mulberry,which we designated MMSR was cloned based on mulberry expressed sequence tags （ESTs）.Sequence analysis showed that the MMSR is 810 base pairs in length,encoding 194 amino acids with a predicted molecular weight of 21.60KD and an isoelectric point of *** expression levels of MMSR at different developmental stages in mulberry leaves and flowers and in different tissues were *** results showed that MMSR transcripts were most abundantly expressed in the bud,and the expression level of the mRNA could be increased significantly under hypothermia,arid,and NaCl stress conditions compared to the normal growth *** is to help understand a molecular basis for signal transduction mechanism underlying the stress response in mulberry.

关键词： Mulberry Methionine sulfoxide reductase (MSR) Gene cloning Sequence analysis Gene expression

Functional analysis of the larval serum protein gene promoter from silkworm,Bombyx mori.

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chinese Science Bulletin 2003年第23期48卷 2611-2615页

作者： TANG Shunming, YI Yongzhu, SHEN Xingjia, ZHANG Zhifang, LI Yiren & HE Jialu Key Laboratory of Silkworm Biotechnology, Ministry of Agriculture, sericultural research institute, chinese academy of agricultural Sci-ences, zhenjiang 212.18, china Correspondence should be addressed to Zhang Zhifang (e-mail: zjsbsri@ ***) Key Laboratory of Silkworm Biotechnology Ministry of Agriculture Sericultural Research Institute Chinese Academy of Agricultural Sciences Zhenjiang 212018 China

The regulation region of larval serum protein gene, Bombyx mori. (BmLSP), consisting of the first intron, the first exon, the central promoter region and 5′-upstream region, is cloned from genomic DNA from the silkworm va-riety of Suju譓inghu. Using PCR and restriction endonu-clease methods, a series of luciferase reporter plasmids, driven by different length of BmLSP promoters, are con-structed. Via the transient expression system in BmN cells, the effects of the regulation elements and foreign insect hor-mones on the BmLSP promoter activity are investigated. The results demonstrate that the promoter activity of BmLSP is 5.8- or 4.4-fold higher than that of BmLSPs whose first in-tron or the element in 5′-upstream region harboring the homologous sequence with the first intron of light-chain fib-roin gene (EHIF) is deleted, respectively, suggesting that both the first intron and EHIF contain the main positive cis-acting elements. However, the inactive mariner transposable ele-ment (MTE) in 5′-upstream region presents a negative effect. Furthermore, the effects of juvenile hormone analogue (JHA) on the BmLSP promoter activity show a typical dose-dependent manner, that is, low concentration treat-ments increase the BmLSP promoter activity and high con-centration treatments decrease it. Meanwhile, insect ecdy-sone (MH) treatments present no significant effect.

关键词：桑蚕血清蛋白基因抑制核酸内切酶法荷尔蒙

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Fine Mapping of I（yellow inhibitor）Gene and Analysis of Candidate Gene in Silkworm（Bombyx mori）

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Fine Mapping of I（yellow inhibitor）Gene and Analysis of Ca...

现代分子植物育种与粮食安全研讨会

作者： Zhang Jun1,Zhang Yuehua2,Wang Xiaoqiang1,Qian Heying2,Xu Anying2* (1 jiangsu University of Science and Technology,zhenjiang 212003,jiangsu,china 2sericultural research institute,chinese academy of agricultural sciences,zhenjiang 212.18,jiangsu.)

The yellow color of the cocoon of the silkworm Bombyx mori is controlled by three genes,Y （Yellow haemolymph）,C （Out-layer yellow cocoon）,and I （Yellow inhibitor）,which are located on linkage groups 2,12,and 9,*** I gene down-regulates the yellow coloring of the haemolymph and controls carotenoid transfer from the midgut epithelium into the *** recently research indicated that the Y gene encodes an intracellular cartenoid-binding protein （CBP） containing a lipid-binding domain known as the steroidogenic acute regulatory protein-related lipid transfer *** the mechanism of I gene remained not to be ***,Li Xia constructed a linkage map of 38.4 cM,and the distance between I gene and the nearest marker S0904 is 7.4 *** learn more about the function and mechanism of the I gene,we performed fine mapping of the I gene in the foundation of preliminary *** to a lack of crossing over in females,reciprocal backcrossed F1 progeny were used for linkage analysis and mapping of the I gene utilizing silkworm strains Ba with white haemolymph （+Y+Y II） and KY with yellow haemolymph （YY+I+I）.We designed SSR,STS and CAPS markers,and finally 1 STS maker and 5 SSR markers were integrated into the initial linkage map by using BC1M[KY×（Ba×KY）] population and Mapmaker 3.0 *** constructed a linkage map of *** I gene located between the marker STS48-1 and S001,and the distance between I gene and the closest marker STS48-1 is at a distane of 0.5 *** four candidate genes may associate with mechanism of the I gene by analyzing the mapping region in NCBI,SilkDB and KaikoBase *** of I gene and studying on the occurring mechanism of I are in progress.

关键词： Fine Mapping Yellow inhibitor gene Candidate Gene Bombyx mori

Sequence analysis and expression of the Calcium-dependent protein kinase gene,MCDPK,in mulberry （Morus L.）

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Sequence analysis and expression of the Calcium-dependent pr...

现代分子植物育种与粮食安全研讨会

作者： Zhang Yinghua1,2,Lin Qiang 3,Liu Zhaoyue1,2,Tong Wei1,2,Fang Rongjun1,2,Liu Li1,2,Zhao Weiguo 1,2 * (1 College of biological and Environmental Engineering,jiangsu University of Science and technology,zhenjiang jiangsu 212.18,china 2 sericultural research institute,chinese academy of agricultural sciences,zhenjiang jiangsu 212.18,china 3 The General Station of Sericulture Technology Extension of Guangxi Zhuang Automomous Region,Nanning 530007,china)

A full-length cDNA sequence coding calcium-dependent protein kinase （CDPK） of mulberry tree,which we designated MCDPK,was cloned based on mulberry expressed sequence tags （ESTs）.Sequence analysis showed that the MCDPK is 2249 base pairs in length,encoding 532 amino acids with a predicted molecular weight of 59.96KD and an isoelectric point of *** expression level of MCDPK at different developmental stages in mulberry leaves and flowers and in different tissues was *** results showed that MCDPK transcripts were most abundantly expressed in the terminal bud,and the expression level of the mRNA could be reduced significantly under the conditions of low temperature and salt stress respectively compared to the normal growth *** research will help to understand the resistance mechanism of functional genes in mulberry in the future.

关键词： Mulberry Calcium-dependent protein kinase gene (CDPK) Gene cloning Gene expression

杆状病毒囊膜糖蛋白gp64基因启动子活性分析(英文)

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生物化学与生物物理学报 2003年第1期35卷 18-26页

作者：周亚竟易咏竹张志芳何家禄张元兴吴祥甫中国农业科学院蚕业研究所、农业部家蚕生物技术重点开放实验室 1中国农业科学院蚕业研究所、农业部家蚕生物技术重点开放实验室华东理工大学生物反应器国家重点实验室中国科学院上海生命科学研究院生物化学与细胞生物学研究所镇江212.18华东理工大学生物反应器国家重点实验室上海200237 华东理工大学在读博士生镇江212.18 上海200237 上海200031

GP6 4是杆状病毒在其发育循环第一时相所产生的芽生型病毒粒子 (buddedvirus ,BV)的特异性囊膜糖蛋白 ,它在病毒入侵细胞过程中具有重要作用。为了探明杆状病毒gp6 4基因的表达调控 ,从所克隆的BmNPV和AcMNPVgp6 4基因ATG上游 437～ 43... 详细信息

GP6 4是杆状病毒在其发育循环第一时相所产生的芽生型病毒粒子 (buddedvirus ,BV)的特异性囊膜糖蛋白 ,它在病毒入侵细胞过程中具有重要作用。为了探明杆状病毒gp6 4基因的表达调控 ,从所克隆的BmNPV和AcMNPVgp6 4基因ATG上游 437～ 439bp启动子的序列分析发现 ,该启动子同时具有早期和晚期转录模体。将所构建的该启动子控制下荧光素酶报告基因 (Luc)的非融合表达质粒分别转染Bm N和Sf 2 1细胞进行瞬间表达分析 ,BmNPVgp6 4启动子能被允许宿主Bm N细胞RNA聚合酶II所识别 ,AcMNPVgp6 4启动子既能被允许宿主Sf 2 1又能被非允许宿主Bm N细胞RNA聚合酶II所识别 ,同时 ,这两个启动子的转录活性在各自的允许宿主细胞中被相应的病毒因子所反式激活 2 .4～ 4倍。将家蚕核多角体病毒同源重复序列 3(BmNPVhomologousregion 3,hr3)克隆到该启动子控制下的Luc报告基因下游 ,用所构建的质粒分别转染细胞 ,瞬间表达分析结果表明 ,BmNPVhr3能分别增强该启动子在Bm N和Sf 2 1细胞中的转录活性 13～ 2 2倍和 70 0 0～ 140 0 0倍以上。同时 ,相应的病毒因子能反式激活插入了hr3的gp6 4启动子在各自允许宿主细胞中转录活性 73～ 78倍 ,这暗示着BmNPVhr3除了具有病毒DNA复制原点和增强子的功能外 ,它在病毒的反式激活过程中?

关键词：杆状病毒 gp64基因启动子同源重复序列3 病毒因子转染

Preliminary Report on A New Germplasm of the Silkworm Mutated in Z Chromosome By Transgenic Method

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Preliminary Report on A New Germplasm of the Silkworm Mutate...

中国生物化学与分子生物学会农业生物化学与分子生物学分会第八次学术研讨会

作者： TANG Shun-Ming~(1,2*) LU Fu-Hao~2 SHEN Xing-Jia~(1,2*) WANG Na~2 ZHAO Qiao-Ling~(1,2) ZHANG Guo-Zheng~(1,2) GUO Xi-Jie~(1,2) (~1 The Key Laboratory of Silkworm and Mulberry Genetic Improvement,Ministry of Agriculture,The sericultural research institute,chinese academy of agricultural sciences,zhenjiang jiangsu 212.18,china ~2 jiangsu University of Science and Technology,zhenjiang jiangsu 212003,china)

In this paper,we reported the new germplasm of silkworm Z by using transgenic *** and molecular genetics analysis results showed that the mutation site is located at Z *** traits around this site was also analyzed and...

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In vitro and in vivo Transient Expression in Insect Cells Mediated by the Cationic Liposome DDAB/DOPE

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In vitro and in vivo Transient Expression in Insect Cells Me...

中国蚕学会第三届青年学术研讨会暨浙江省第二届青年学术论坛—蚕桑分论坛

作者： Qing Li Xiao, Ya Jing Zhou, Zhi Fang Zhang, Jia Lu HeKey Laboratory of Silkworm Biotechnology, Ministry of Agriculture, sericultural research institute, chinese academy of agricultural sciences, zhenjiang, jiangsu, 212.18, P. R. china

Cationlc liposomes complexed with DNA have been extensively utilized for the delivery of reporter ortherapeutic genes both in culture and Jn *** investigated and determined the optimum conditions of a cationic liposome,composed of dimethyldioctadecylammonium bromide(DDAB)and deoleoyl phosphatidylethanolamine(DOPE),mediated areporter plasmld expressing luciferase into insect cell lines(Sf-21 and Bm-N)and silkworm *** the datademonstrated that Bombyx mori nuclear polyhedrosis virus(BmNPV)genomic DNA(128 kb)was successfully transfectedinto Bm-5 cells using this *** results suggest that DDAB/DOPE liposome will be useful as delivery agents forgene transfer to insecet cells both in vitro and in vivo.

关键词： Cationic liposome transient expression insect cells silkworm larvae