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2005年05月28日

【期刊论文】siRNA介导的家蚕ABC转运蛋白相关基因的干涉研究

夏庆友, 王根洪, 刘春, 侯勇, 李娟, 赵萍

,-0001,():

-1年11月30日

摘要

由siRNA(short interfering RNAs)介导的RNAi作为一种工具已被广泛用于基因功能研究中,并有望在不久的将来成为防治生物体疾病的强大工具。本研究通过显微注射,在家蚕中首次以siRNA介导的方式,对家蚕ABC 转运蛋白(ATP Binding Cassette Transporter)系列相关基因(Bmwh1,Bmwh2,Bmwh3)进行了siRNA干涉研究,在部分基因中得到了高效特异的干涉现象。比较分析结果初步证实siRNA正义链3’碱基错配可以增强siRNA的干涉效果。同时,实验显示了siRNA较之于dsRNA介导的RNAi具有较强的优势,表明siRNA介导的RNAi可作为基因功能分析和未来的基因操作手段在家蚕中进行应用。

siRNA, RNAi, ABC, 转运蛋白, 显微注射

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2005年05月28日

【期刊论文】EST-based Profiling and Comparison of Gene Expression in the Silkworm Fat Body during Metamorphosis

夏庆友, Dao-Jun Cheng, Qing-You Xia *, Ping Zhao, Zi-Long Wang, Han-Fu Xu, Guan-Rong Li, Cheng Lu, and Zhong-Huai Xiang

,-0001,():

-1年11月30日

摘要

The fat body plays key roles in metabolism and sustenance of growth throughout the life of a silkworm, and thus represents a model tissue for studying development and metamorphosis. Analysis of 18480 ESTs derived from larval and pupal fat body cDNA libraries allowed characterization of gene expression patterns in the fat body during metamorphosis. By Phrap assembling, 6814 estimated transcription units (TUs) were generated, 2673 of which contained 60% of the total ESTs and represented 1366 distinctive known genes. These genes were classified into 14 categories based on their known or putative functions, and most were found to be involved in metabolism and cell defense. Further comparative analysis of the expression profiles revealed that the gene expression patterns in the larval fat body strikingly differed from those in the pupal fat body. From this, we identified a subset of genes potentially associated with metamorphic events such as the formation or destruction of specific tissues, and simultaneously confirmed the stage-specific expression patterns of several identified genes using RT-PCR examination. This work will provide a valuable resource for studying regulatory mechanisms associated with fat body developmental changes and silkworm metamorphosis.

Silkworm, Fat body, EST, Trans, c, r, i, p, t, ome, Alternation, Metamorphosis

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2005年05月28日

【期刊论文】家蚕不同品种与个体的血液胰凝乳蛋白酶抑制剂差异研究

夏庆友, 赵萍, 藤井博, 伴野丰

蚕业科学,2003,29(4):349~354,-0001,():

-1年11月30日

摘要

胰凝乳蛋白酶抑制剂(Chymotrypsin inhibitor,CI)是家蚕血液中主要的蛋白酶抑制因子,为其正常生长发育所不可缺少,研究a对阐明家蚕正常生理以及害虫防治等都具有重要意义。采用pH8.3、4.0聚丙烯酰胺凝胶电泳和CI活性染色法,对6个家蚕品种血液中的蛋白质含量和CI活性进行了测定,并对各品种CI活性的个体差异性进行了分析。结果表明:结合pH 8.3和pH4.O的电泳条件,可检测出Ict-D,Ict-E,Ict-A,Ict-B,Ict-H基因所支配的15种CI;6个品种中有3个具有CI个体差异性,而且Ict-E和Ict-B在有的品种或个体无活性,为非必需种类。

家蚕, 血液, 胰凝乳蛋白酶抑制剂, 比活性, 个体差异

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2005年05月28日

【期刊论文】Mining single nucleotide polymorphisms from EST data of silkworm, Bombyx mori, inbred strain Dazao

夏庆友, Ting-Cai Cheng, Qing-You Xia *, Ji-Feng Qian, Chun Liu, Ying Lin, Xing-Fu Zha, Zhong-Huai Xiang

Insect Biochemistry and Molecular Biology 34(2004)523-530,-0001,():

-1年11月30日

摘要

We made use of 81,635 expressed sequence tags (ESTs) derived from 12 different cDNA libraries of the silkworm, Bombyx mori, inbred strain Dazao (P50), to identify high-quality candidate single nucleotide polymorphisms (SNPs). By PHRAP assembling, 12,980 contigs containing 11,537 contigs assembled by more than one read were obtained, and 101 candidate SNPs and 27 single base insertions/deletions were identified from 117 contigs assembled from 1576 high-quality reads base-called with PHRED and screened on the basis of the neighborhood quality standard (NQS). Simultaneously, we also predicted 40 SNPs in coding regions (cSNPs), of which 26 were predicted to lead to amino acid non-synonymous variations and 14 synonymous substitutions. Also, the 1.66:1 ratio of transition/transversion is different from that of other insects. As the first SNP analysis of a Lepidoptera, B. mori, the single nucleotide polymorphic density is estimated to be 1:3 103 by sequence diversity. This analysis shows that expressed sequences from multiple libraries may provide an abundant source of comparative reads to mine for cSNPs from the silkworm genome.

Single nucleotide polymorphism, cSNP, EST, Leopidoptera, Bombyx mori

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2005年05月28日

【期刊论文】家蚕母性基因的表达序列标签分析

夏庆友, 邱咏梅, 夏庆友*, 程道军, 沈以红, 刘春, 林英, 查幸福, 向仲怀

昆虫学报,2004,47(2):159-165,-0001,():

-1年11月30日

摘要

为了研究家蚕Bombyx mori卵中储存的母性遗传信息,以未受精的肾脏型和正常型蚕卵为材料构建了cDNA文库,并对其随机测序,共获得391条表达序列标签(EST)序列,经拼接后得到374条非重复序列。生物信息学分析发现,172个非重复序列与国际数据库GenBank和silkbase中的已知基因具有较高的相似性。除3个已知母性基因外,其余主要与DNA复制、能量代谢、信号转导、转录、蛋白质结构以及胚胎发育中个体形成等相关:该结果可为进一步研究家蚕胚胎早期发育的调控提供基础数据。

家蚕, 表达序列标签, 未受精卵, 胚胎发育, 母性基因

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    西南农业大学,重庆

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