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Study of Different Expression Genes in the Near-isogenic Lines of Dominant Endurance to Fluoride(Def)gene of the Silkworm, Bombyx Mori

Author: WangXiaoQiang
Tutor: XuAnYing
School: Jiangsu University of Science and Technology
Course: Special Economic Animal Feeding
Keywords: Bombyx mori Def gene near-isogenic line SSH 2-DE Real-time PCR
CLC: S882.1
Type: Master's thesis
Year: 2011
Downloads: 8
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Abstract

China is the origin of sericulture in the world. The silk industry in our country have over 5,000 years of history, includeing many sections, such as planting mulberry, feeding silkworms, reeling silk from cocoons, knitting the silk and processing garment. Sericulture is a very important part of Chinese agriculture. Silk industry is a complete system composed of agriculture, industry and commerce. Silk exportation holds an important position in Chinese national economy.The fluoride tolerance is the one of important economy traits in silkworm. Mulberry leaves are the major source of food for silkworm.It caused a large number of deaths to the silkworm when mulberry leaves were polluted by fluoride. This could cause serious harm to the sericulture. So it is very important to do works such as studying poisoning mechanism of fluoride in silkworm, searching for dominant endurance to fluoride(Def)gene of silkworm, breeding new new varieties of silkworm which are endurable to fluoride, map-based cloning of Def gene.Silkworm near-isogenic lines(NILs)of dominant endurance to fluoride(Def)gene were constructed using silkworm strain 733 xin (sensitive to fluoride) and T6 (endurable to fluoride). An SSH (suppression subtractive hybridization) cDNA library was constructed between individuals which were sensitive to fluoride and individuals which were endurable to fluoride of BC11 progeny using PCR-selectTM cDNA Subtraction Kit. Plasmids were extracted from 153 positive clones randomly selected from the library and then sequenced and assembled using CAP3 software. 19contigs and 48 unigenes were obtained. Blastx searches and homology analysis with existing expressed sequence tags(ESTs)revealed that these genes were involved in many biological processes such as substance transportation, energy metabolism, gene transcription, protein synthesis or degradation, protein processing, signal transduction, organismal immunologic defence and formation of cellular construction etc. Real-time qPCR analysis confirmed that some genes, such as phosphate transport protein, ATP synthase, vacuolar-type H+-ATPase and major allergen protein significantly up-regulated in midguts, malpighian tubules, fat body of individuals which were endurable to fluoride. Our results provide useful informations for further studies on molecular mechanism of endurable to fluoride of silkworm.2-DE and mass spectroscopy Analysis was done at the individual protein expressed difference between individuals which were sensitive to fluoride and individuals which were endurable to fluoride from the near-isogonic lines of Def gene in silkworm. 5 different protein spots that can trusted were gotten. There are 2 protein spots that are relative to energy metabolize and immunity, and we can’t confirm the function of the rest 3 protein spots. This study of this period may lay a foundation for deep researches on function of protein in future.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Sericulture > Genetic breeding and breeding of the silkworm > Silkworm germplasm and varieties save
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