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国家重点基础研究发展计划(2004CB117404)

作品数:11 被引量:168H指数:7
相关作者:梁利群雷清泉常玉梅孙效文高俊生更多>>
相关机构:哈尔滨理工大学中国水产科学研究院黑龙江水产研究所大连水产学院更多>>
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11 条 记 录,以下是 1-10
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与鲤鱼抗寒性状相关的RAPD分子标记的筛选及其克隆被引量:13
2006年
以黑龙江鲤(Cyprinus carpio haematopterusTemminck et Schlegel)、荷包红鲤抗寒品系(Cyprinus carpiovar.wana-nensis)、荷包红鲤(Cyprinus carpiovar.wananensis)、柏氏鲤(Cyprinus pellegniniTchang)以及荷包红鲤抗寒品系和柏氏鲤的杂交F2的DNA样品为材料,用200个随机引物对其进行PCR扩增,获得了1个与鲤鱼抗寒性状相关的分子标记RAG20,并在F2代分离个体中得到验证。至此,本实验室共获得10个与鲤鱼抗寒性状相关的分子标记,更加证明抗寒性状是数量性状(QTL)受微效多基因控制。此外,将其中4个标记从琼脂糖凝胶上进行回收,并与pMD18-T vector连接,将其转入感受态大肠杆菌E.coliDH 5α,对克隆片段进行序列分析,旨为根据每一序列重新设计引物,将RAPD标记转化为稳定的SCAR标记。
梁利群高俊生李绍戊孙效文雷清泉
关键词:鲤鱼抗寒SCARRAPD分子标记
Genome evolution trend of common carp (Cyprinus carpio L.) as revealed by the analysis of microsatellite loci in a gynogentic family被引量:7
2008年
Genome evolution arises from two main ways of duplication and reduction. Fish specific genome duplication (FSGD) may have occurred before the radiation of the teleosts. Common carp (Cyprinus carpio L.) has been considered to be a tetraploid species, because of its chromosome numbers (2n=100) and its high DNA content. Using 69 microsatellite primer pairs, the variations were studied to better understand the genome evolution (genome duplication and diploidization) of common carp from a gynogenetic family. About 48% of primer pairs were estimated to amplify duplicates based on the number of PCR amplification per individual. Segregation patterns in the family suggested a partially duplicated genome structure and disomic inheritance. This indicates that the common carp is tetraploid and polyploidy occurred by allotetraploidy. Two primer pairs (HLJ021 and HLJ332) were estimated to amplify reduction based on the number of PCR amplification per individual. One allele in HLJ002 locus and HLJ332 locus was clearly lost in the gynogenetic family and the same as in six wild populations. Segregation patterns in the family suggested a partially diplodization genome structure. A hypothesis transition (dynamic) and equilibrium (static) were proposed to explain the common carp genome evolution between genome duplication and diploidization.
Yah ZhangLiqun LiangPeng JiangDayu LiCuiyun LuXiaowen Sun
鲤抗寒性状的RAPD标记转化为SCAR标记的研究被引量:10
2007年
以黑龙江鲤Cyprinus carpio haematopterus、荷包红鲤抗寒品系、荷包红鲤Cyprinus carpio var.、柏氏鲤Cyprinus pellegnini pellegnini以及荷包红鲤抗寒品系(父本)与柏氏鲤(母本)杂交F2的DNA样品为材料,用300个随机引物对其进行PCR扩增,获得2个与鲤抗寒性状相关的分子标记AL04986和AR02788。回收、纯化这两个RAPD标记,连接到pMD18-T载体中,转化DH5α大肠杆菌感受态细胞,并对插入片段进行测序。根据测序结果,设计了3对SCAR引物SCAL04L/SCAL04R1、SCAL04L/SCAL04R2和SCAR02L/SCAR02R,其中SCAR02L/SCAR02R这对引物可以成功的反映出杂交F2两个群体在抗寒能力上的差异,适宜的退火温度为57℃,将此标记命名为SCAR02788。根据AL04986的测序结果设计的两对SCAR引物并没有反映出这种差异。SCAR02L/SCAR02R标记可以用作鲤抗寒基因分子辅助选择的依据。
高俊生孙效文梁利群
关键词:抗寒RAPD标记SCAR标记分子标记
低温对鲤鱼的5个基因在不同组织中表达量的影响被引量:8
2008年
本研究应用实时荧光定量PCR技术对抑制性消减杂交所得到的鲤鱼不同温度下差异表达的5个基因做进一步的研究。结果表明:水温由16℃降至4℃(骤降点)的过程中,在鲤鱼的肝、肠组织中,LKE-25基因的表达量呈下降趋势,而LKE-62基因则呈上升趋势;在肾脏组织中,LKE-48-1、LKE-48-2和LKE-59基因的表达量呈上升趋势,且随着4℃维持时间的延长,其表达量逐渐下降。LKE-25和LKE-59基因在4℃同一尾鱼的心脏、脑、肝、脾、肾、肠组织中的表达量上有很大差异,其中LKE-25基因在心、LKE-59基因在肾组织中的表达量最高。本实验所研究的5个基因在不同温度、不同组织表达量的显著差异,证明了这些基因与水温相关且有组织特异性。
李勇丁雷李言侯宁常玉梅梁利群雷清泉
关键词:鲤鱼实时荧光定量PCR
筛选与鲤鱼抗寒性状相关的微卫星分子标记被引量:16
2008年
为了寻找鲤鱼中与抗寒性状相关的DNA序列,采用142个微卫星引物,对荷包红鲤抗寒品系、柏氏鲤及其杂交F2代中的青灰色抗寒与青灰色不抗寒个体的混合基因组DNA进行分析.经过710次微卫星分子标记检测,首次获得两个(引物HLJ578,HLJ580)与抗寒性状相关的微卫星分子标记.利用这些微卫星分子标记进一步对27个抗寒与不抗寒个体的基因组DNA进行检测,验证其准确性.结果表明:鲤鱼的抗寒性状是数量性状,且受微效多基因调控;与鲤鱼抗寒相关分子标记DNA序列的拷贝数在鲤鱼抗寒与不抗寒个体中存在差异,这些与抗寒性状相关的DNA序列的拷贝数目的多少将最终影响与抗寒性状有关的蛋白质表达量的多少.
潘贤梁利群雷清泉
关键词:鲤鱼微卫星分子标记
低温胁迫对鲤血液学和血清生化指标的影响被引量:40
2006年
A total of 85 interspecific hybrid F2 (Cyprinus carpio var.wuyuanensis ×Cyprinus pellegrini pellegrini) were cooled to specific temperatures and held at those temperatures over a maximum of 4 days in a water-recycled and temperature-controlled aquarium inside.As a result,the blood homeostasis of experimental fish changed violently as acute temperature changed from 16 ℃ to 10 ℃ and 4 ℃ at a rate of 1 ℃·h-1 according to the data we collected.Whole blood pH,also called extracellular pH(pHe) were very sensitive to temperature changes,where there was a significant difference between 10 ℃(7.41) and 16 ℃(7.17)(P<0.01),compared to other values of hematology and serum chemistry.When the water temperature was continually decreased to an extreme temperature of 4 ℃,the content of Na+ of serum decreased remarkably in comparison with that of 10 ℃ and 16 ℃,which was 85.2 mmol·L-1,113.3 mmol·L-1 and 118.7 mmol·L-1,respectively.The values of hematology and serum chemistry also altered in gentle temperature changes of(10±2) ℃ and(4±2) ℃.Most values of serum chemistry and pH changed significantly,whereas the values of blood plasma changed slightly.pH was up slowly in 4 days at(10±2) ℃ and down slowly in 3 days at(4±2) ℃.A variety of values of serum chemistry changed remarkably both at(10±2) ℃ and(10±2) ℃,but the values of TP,TG and ALB only changed significantly at(4±2) ℃.These results distinguished at least two mechanisms involved in cold-induced stress in hybrid F2.Cold-induced pH changes resulted in other values altered.What’s more,pH correlated negatively with water temperature above 10 ℃,and the content of Na+.We also found that gentle temperature changes will be physiologically compensated for on day one at(10±2) ℃ and on day 2 at(4±2) ℃ in hybrid F2.
常玉梅匡友谊曹鼎臣梁利群孙效文雷清泉
关键词:胁迫
A gene family-based method for interspecies comparisons of sequencing-based transcriptomes and its use in environmental adaptation analysis被引量:1
2010年
We describe a new method for sequencing-based cross-species transcriptome comparisons and define a new metric for evaluating gene expression across species using protein-coding families as units of comparison. Using this measure transcriptomes from different species were evaluated by mapping them to gene families and integrating the mapping results with expression data. Statistical tests were applied to the transcriptome evaluation results to identify differentially expressed families. A Perl program named Pro-Diff was compiled to im- plement this method. To evaluate the method and provide an example of its use, two liver EST transcriptomes from two closely related fish that live in different temperature zones were compared. One EST library was from a recent sequencing project of Dissosticus maw- soni, a fish that lives in cold Antarctic sea waters, while the other was newly sequenced data (available at: http://www.fishgenome.org/ polarbank/) from Notothenia angustata, a species that lives in temperate near-shore water of southern New Zealand. Results from the com- parison were consistent with results inferred from phenotype differences and also with our previously published Gene Ontology-based method. The Pro-Diffprogram and operation manual can be downloaded from: http://www.fishgenome.org/download/Prodiff.rar.
Zuozhou ChenHua YeLonghai ZhouChi-Hing C. ChengLiangbiao Chen
关键词:EST
抑制消减杂交技术在鲤鱼抗寒研究中的应用被引量:22
2006年
用抑制性消减杂交技术(SSH)研究与鲤鱼(Cyprinus carpio)抗寒性状相关的基因。通过对荷包红鲤抗寒品系(Cyprinus carpio wuyanensis Tchang)和柏氏鲤(Cyprinus pellegnini Tchang)杂交F2代进行降温诱导实验,获得不同温度下的实验材料。利用抑制消减杂交技术构建了一个低温下差异表达基因的消减cDNA文库,共含有2000个克降。对其叶1480个克隆进行斑点杂交筛选,共得到60个阳性克隆,选取其中29个克隆测序。通过序列分析比对,13个克隆与已知基因序列高度同源,同源性为85%-98%;另外的13个克隆为未知新基因序列。在13个同源性较高的克隆序列巾,有brevican基因、AMP—acid连接酶、CFL2基因、催产素基因、主要组织兼容复合体(MHC)Ⅱβ链、锌指蛋白、细胞色素氧化酶、EPD-1基因、透明质酸结合蛋白多糖类、核受体/类视色素信号调节器等,其中9个基因上调表达,另有4个则抑制表达。这些基因的获得可为研究鱼类抗寒性状以及从分子水平上阐明其机制提供重要依据。
梁利群李绍戊常玉梅高俊生孙效文雷清泉
关键词:抑制消减杂交鲤鱼
鲤NADH泛醌氧化还原酶亚基3基因的克隆及低温适应相关性分析被引量:3
2009年
根据鲤低温下特异表达的基因(片段)序列设计引物,采用2轮菌落PCR方法对鲤脑全长cDNA文库中的克隆进行筛选,确定了基因序列152在文库中的位置,经生物信息学比对分析确定其为NADH泛醌氧化还原酶亚基3基因.这一基因在低温下的特异表达,对于低温下鱼类机体的供能,起着重要作用.结果还表明,NADH泛醌氧化还原酶亚基3基因在鲤氧化呼吸链电子传递系统中与低温适应性状在一定程度上存在相关性.
梁利群常玉梅邹庆薇雷清泉
关键词:克隆
鲤鱼体长性状的QTL定位及其遗传效应分析被引量:69
2007年
以大头鲤/荷包红鲤抗寒品系的重组自交系群体及其遗传连锁图谱,利用Windows Map Manager 2.0的标记回归法进行QTL单标记定位分析和复合区间作图法进行QTL区间检测,通过置换实验(1 000次重复)确定连锁群显著性水平阈值。在体长性状的标记回归研究中,共7个标记达到显著水平(P<0.01),对性状的贡献率为14.00%~27.00%,其中3个标记达到极显著水平(P<0.001)。HLJ534,HLJ319,HLJ370座位可能与影响鲤鱼体长性状的主效基因连锁。在体长性状的QTL区间定位研究中,共6个QTL达到连锁群显著水平(P=0.05),对性状的贡献率为11.33.%~23.12%,其中2个达到连锁群极显著性水平(P=0.01),它们的加性效应方向并不一致。HLJ190-HLJ497区间和HLJ479-HLJ483区间是影响鲤鱼体长性状的主效QTL区间。
张研梁利群常玉梅侯宁鲁翠云孙效文
关键词:鲤鱼微卫星体长
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