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国家重点基础研究发展计划(2006CB102101)

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藏鸡和两种低地鸡线粒体全基因组序列的测定和比对被引量:4
2007年
藏鸡生活在青藏高原,它对高原低氧具有一定的遗传适应能力.丝羽乌骨鸡和寿光鸡都是低地鸡种,缺乏对低氧环境的遗传适应能力.鸡的线粒体基因组总长16785bp,共编码37个基因,这些基因的编码产物均与线粒体的呼吸作用和氧化磷酸化相关,它们的突变均有可能影响线粒体的功能,因此均可被选为藏鸡低氧遗传适应的候选基因.测定比较藏鸡和低地鸡的线粒体基因组,可以为进一步研究藏鸡的低氧遗传适应机理提供线索.本研究测定和比较了藏鸡、丝羽乌骨鸡和寿光鸡的线粒体全基因组序列,发现藏鸡线粒体基因组总长为16784—16786bp,丝羽乌骨鸡为16785bp,寿光鸡为16784bp;比对后共发现突变(包括单核苷酸多态性(SNP)、单碱基缺失和插入)120个,其中tRNA基因突变4个,rRNA基因突变10个,D—LOOP区突变39个,基因间隔区突变1个,编码蛋白质亚基的碱基突变66个(包括同义突变48个,错义突变18个).
鲍海港赵春江李俊英吴常信
关键词:藏鸡线粒体基因组低氧
中国美利奴羊IGFBP-3基因内含子4区的PCR-RFLP多态性分析被引量:4
2010年
本研究针对在绵羊IGFBP-3基因内含子4区新发现的1个SNP位点(C218T),建立了PCR-RFLP的基因分型方法。利用该方法对353只中国美利奴羊(新疆军垦型)进行了多态性检测,结果显示,有CC(249bp)、CT(249/216/33bp)和TT(216/33bp)3种基因型,基因型频率分别为0.52、0.41和0.07;有C、T2种等位基因,等位基因频率分别为0.72、0.28。多重比较结果表明,不同基因型对中国美利奴部分羊毛生产性能有一定的影响,CC基因型个体的毛丛长度显著短于TT基因型个体(P<0.05),其毛囊密度显著高于CT和TT基因型个体(P<0.05)。
沈敏王文君杨永林马春萍刘守仁李宁
关键词:胰岛素样生长因子结合蛋白3单核苷酸多态性羊毛性状中国美利奴羊
线粒体ND5基因的变异对藏鸡胚胎脑线粒体呼吸功能的影响
引言我们以前的研究表明藏鸡和低地鸡胚胎在脑线粒体呼吸功能上的差异可能与NADH脱氢酶复合体有关。对藏鸡线粒体全基因组序列的测定结果表明,在9个藏鸡特有的错义突变中,有7个是在NADH脱氢酶复合
鲍海港赵春江吴常信
关键词:藏鸡胚胎线粒体呼吸功能
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猪CACNA1S基因部分序列的克隆、测序及SNP检测被引量:3
2008年
CACNA1S是钙离子通道主效亚基α1亚单位的编码基因,该基因突变会导致人(Homo sapiens)低钾性周期性麻痹和恶性高温综合征.目前CACNA1S基因的研究主要集中在人和模式动物上,而在家畜中的研究少见报道.本研究以金华猪(115头)、皮特兰猪(30头)、金华猪与皮特兰猪杂交产生的金皮F2代杂种猪(126头)、大约克猪(23头)为研究对象,根据人CACNA1S基因序列设计引物,对猪基因组DNA进行PCR扩增、克隆测序,并与人相应序列作同源性比较,然后采用PCR-SSCP技术检测序列中可能存在的单核苷酸多态(single nucleotide polymorphism,SNP)位点,并对有合适内切酶存在的突变点应用PCR-RFLP技术进行验证.结果表明:(ⅰ)用6对引物对猪基因组DNA进行扩增,共克隆得到猪CACNA1S基因约5211bp的DNA序列,其中外显子区域,猪与人同源性为82.6%,序列已递交GeneBank收录(登录号DQ767693);(ⅱ)在克隆得到的DNA序列中,共检测到57个单核苷酸突变点,其中外显子区域存在24个;(ⅲ)突变位点的PCR-RFLP验证与PCR-SSCP检测结果一致,经与GenBank公布的猪CACNA1S基因EST小片段(Bx914582,Bx666997)比较,相应长度核酸序列内本实验检测到的11个SNP位点中,有8个位点的碱基突变与2个EST片段间存在的碱基差异相同.
方晓敏徐宁迎任守文
关键词:PCR-SSCPPCR-RFLP
Study on Tibetan Chicken embryonic adaptability to chronic hypoxia by revealing differential gene expression in heart tissue被引量:5
2009年
Oxygen concentration is essential for appropriate metabolism.Hypoxia can exert a significant impact on physiological alteration of the cell and organism.Tibetan Chicken(Gallus gallus) is a Chinese indigenous breed inhabiting in Tibetan areas,which is also a chicken breed living at high altitude for the longest time in the world.It has developed an adaptive mechanism to hypoxia,which is demonstrated by that Tibetan Chicken has much higher hatchability than low-land chicken breeds in high-altitude areas of Tibet.In the present study,Tibetan Chicken fertilized full sib eggs were incubated up to Hamburger-Hamilton stage 43 under 13% and 21% oxygen concentration,respectively.Shouguang Chicken and Dwarf Recessive White Chicken were used as control groups.The hearts in all of the 3 chicken breeds under hypoxic and normoxic conditions were isolated and hybridized to Genechip Chicken Genome Array to study molecular mechanisms underlying the adaptation to high altitude of Tibetan Chicken.As a result,50 transcripts highly expressed in hypoxia are screened out.Among up-regulated genes,some are involved in the gene ontology(GO) such as cell growth,cell difference,muscle contraction and signal transduction.However,the expression levels of 21 transcripts are lower in hypoxia than those in normoxia.Some down-regulated genes take part in cell communication,ion transport,protein amino acid phosphorylation and signal transduction.Interestingly,gene enrichment analyses of these differential gene expressions are mainly associated with immune system response and ion channel activity in response to stimulus.Moreover,the transcriptional expression profiles analyzed by hierarchical clustering and CPP-SOM software in all of the 3 different chicken breeds revealed that Tibetan Chicken is much closely related to Shouguang Chicken rather than Dwarf Recessive White Chicken.In addition,12 transcripts of Tibetan Chicken breed-specific expressed genes were identified,which seem to result in a more effective and efficient induction of energy dema
LI Mei & ZHAO ChunJiang College of Animal Science and Technology,China Agricultural University,Beijing 100193,China
关键词:GENECHICKENGENE
猪泛素-B基因的克隆及组织表达谱分析被引量:3
2008年
从人泛素-B(Ubiquitin B,UbB)基因mRNA序列出发,在猪的ESTs库中进行同源性搜索,通过电子克隆和进一步RT-PCR方法从猪肌肉组织总RNA中扩增出泛素-B基因部分cDNA序列(GenBank登录号:EF688558),长894bp,其中在89~778bp处包含一个完整的开放阅读框,编码229个氨基酸。猪泛素-B基因由2个外显子和1个内含子构成(GenBank登录号:EF688559),内含子长657bp,符合GT-AG规则。序列分析结果表明:猪泛素-B基因与已报道的人、黑猩猩、小家鼠、大鼠、鸡等物种的泛素-B基因高度同源,同源性分别为92%、91%、91%、91%、89%,氨基酸序列同源性为100%。半定量RT-PCR结果表明:泛素-B基因在150日龄肥育猪心脏、肝脏、肾脏、小脑中的表达量较高,脾脏、肺脏、胃、膀胱、背最长肌、大脑次之,脂肪和下丘脑中的表达量最低,其在蓝塘与长白猪肝脏、脾脏、肺脏、肾脏、胃、膀胱、背最长肌等7个组织中的表达量存在差异。
谢水华李加琪陈瑶生张豪王翀梅盈洁莫德林李泽月
关键词:基因表达
利用PCR-SacⅡ-RFLP技术检测绵羊IGFBP-3基因多态性的研究被引量:3
2010年
【目的】将IGFBP-3基因作为候选基因,寻找与绵羊羊毛品质等经济性状相关的SNP位点。【方法】利用PCR-SSCP和DNA测序快速筛查SNP位点;在此基础上建立PCR-RFLP检测方法,分析该位点在不同品种绵羊中的基因型和等位基因分布情况;通过多重比较分析不同基因型与中国美利奴羊部分羊毛性状的关联性。【结果】以绵羊基因组DNA为模板扩增得到了249 bp的特异条带,PCR-SSCP及测序分析显示在该序列的140碱基处发生了G/T突变,导致SacⅡ酶切位点消失。经PCR-SacⅡ-RFLP检测,哈萨克羊和中国美利奴羊细毛羊得到三种基因型:TT(249 bp)、GG(139 bp/110 bp)和TG(249 bp/139 bp/110 bp);杜泊羊中出现GG和TG两种基因型;湖羊、小尾寒羊以及萨福克羊中只有GG一种基因型。关联性分析表明不同基因型与部分羊毛性状没有明显的相关性。【结论】在绵羊IGFBP-3基因嫩含子4区发现一个新的SNP位点,玻建立PCR-SacⅡ-RFLP检测方法;该位点不同等位基因和基因型在不同绵羊品种中的分布具有一定规律性;不同基因型与中国美利奴细毛羊部分羊毛性状中间没有明显的相关性。
沈敏王文君杨永林甘尚权马春萍何其宏张永胜李宁刘守仁
关键词:胰岛素样生长因子结合蛋白3单核苷酸多态性羊毛性状绵羊
Sequencing and alignment of mitochondrial genomes of Tibetan chicken and two lowland chicken breeds被引量:2
2008年
Tibetan chicken lives in high-altitude area and has adapted well to hypoxia genetically. Shouguang chicken and Silky chicken are both lowland chicken breeds. In the present study, the complete mito-chondrial genome sequences of the three chicken breeds were all sequenced. The results showed that the mitochondrial DNAs (mtDNAs) of Shouguang chicken and Silky chicken consist of 16784 bp and 16785 bp respectively, and Tibetan chicken mitochondrial genome varies from 16784 bp to 16786 bp. After sequence analysis, 120 mutations, including 4 single nucleotide polymorphisms (SNPs) in tRNA genes, 9 SNPs and 1 insertion in rRNA genes, 38 SNPs and 1 deletion in D-LOOP, 66 SNPs in pro-tein-coding genes, were found. This work will provide clues for the future study on the association between mitochondrial genes and the adaptation to hypoxia.
BAO HaiGang, ZHAO ChunJiang, LI JunYing & WU ChangXin College of Animal Science and Technology, China Agricultural University, Beijing 100094, China
关键词:TIBETANCHICKENLOWLANDCHICKENMITOCHONDRIALHYPOXIA
Cloning, sequencing and identification of single nu-cleotide polymorphisms of partial sequence on the porcine CACNA1S gene被引量:2
2008年
CACNA1S gene encodes the α1 subunit of the calcium channel. The mutation of CACNA1S gene can cause hypokalemic periodic paralysis (HypoKPP) and maliglant hyperthermia synarome (MHS) in hu-man beings. Current research on CACNA1S was mainly in human being and model animal, but rarely in livestock and poultry. In this study, Yorkshire pigs (23), Pietrain pigs (30), Jinhua pigs (115) and the second generation (126) of crossbred of Jinhua and Pietrain were used. Primers were designed ac-cording to the sequence of human CACNA1S gene and PCR was carried out using pig genome DNA. PCR products were sequenced and compared with that of human, and then single nucleotide poly-morphisms (SNPs) were investigated by PCR-SSCP, while PCR-RFLP tests were performed to validate the mutations. Results indicated: (1) the 5211 bp DNA fragments of porcine CACNA1S gene were ac-quired (GenBank accession number: DQ767693 ) and the identity of the exon region was 82.6% be-tween human and pig; (2) fifty-seven mutations were found within the cloned sequences, among which 24 were in exon region; (3) the results of PCR-RFLP were in accordance with that of PCR-SSCP. Ac-cording to the EST of porcine CACNA1S gene published in GenBank (Bx914582, Bx666997), 8 of the 11 SNPs identified in the present study were consistent with the base difference between two EST frag-ments.
FANG XiaoMin1,2, XU NingYing2 & REN ShouWen1 1 Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
关键词:SUBUNITSINGLENUCLEOTIDE
IGFBP-3基因多态性及其与中国美利奴羊部分羊毛性状的关联性分析被引量:15
2008年
采用PCR-SSCP方法对中国美利奴羊和哈萨克羊中IGFBP-3基因的多态性进行了检测,并对不同基因型与中国美利奴羊部分羊毛性状间的关联性进行了分析。结果在位于内含子1区的一段178bp的扩增产物经SSCP分析后出现了3种基因型,基因型AA、AB和BB及等位基因A、B在中国美利奴羊中的频率分别为0.70、0.24、0.06和0.82、0.18;在哈萨克羊中的频率分别为0.87、0.13、0.00和0.93、0.07。序列分析发现:在该序列的122位碱基表现多态性(g.122G>T)。所研究的两个群体在该位点上均处于Hardy-Weinberg不平衡状态(P<0.01)。不同基因型对部分羊毛性状有一定的影响:不同基因型个体在剪毛后体重和净毛率上没有明显差异。AA、AB及BB基因型个体的羊毛伸直长度逐渐变短,其中AA与AB基因型之间差异极显著(P<0.01)。AA型个体的剪毛量和羊毛密度要明显低于AB型(P<0.01)和BB型个体(P<0.05);羊毛纤维直径则明显高于AB型(P<0.01)和BB型(P<0.05)个体。
沈敏王文君杨永林甘尚权何其宏张永胜王建华马春萍刘正山刘守仁李宁
关键词:中国美利奴羊IGFBP-3基因多态性羊毛性状
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