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1.
根据Sundick等发表的鸡白细胞介素-2(IL-2)基因序列设计一对特异性引物,从伴刀豆球蛋白(ConA)体外激活的脾淋巴细胞中提取mRNA,通过RT-PCR方法获得艾维茵商品肉用鸡IL-2 cDNA基因。将其连接到测序载体pGEM-T Easy Vector,测序后用Pcgene和DNAstar软件分析。结果显示,艾维茵商品肉用鸡IL-2基因长734bp,编码一由143个氨基酸组成的前体蛋白,与来自GenBank的另外3个品系(Kestral leghorn,Obese和SC)鸡IL-2比较,其核苷酸和氨基酸序列同源性为98.9%-100%和97.2%-100%。二级结构预测表明,艾维茵商品肉用鸡IL-2N端存在一条由22个氨基酸组成的前导肽及4个关胱氨酸形成的链内二硫键,并存在类似人IL-2的4个α螺旋,这些结构很可能在维持IL-2生物学活性中发挥重要作用。基因系统进化树分析表明,艾维茵商品肉用鸡与SC鸡IL-2的新缘关系较近,与火鸡、Kestrel leghorn和Obese鸡IL-2的亲缘关系较远。  相似文献   

2.
3.
鸡IL-2基因的克隆及GST-chIL2融合蛋白的表达   总被引:1,自引:1,他引:0  
根据Sundick等发表的鸡IL-2基因(chIL2)序列设计合成特异性引物,用RT-PCR从ConA诱导的鸡脾淋巴细胞扩增出450 bp的目的片段,酶切鉴定及序列测定结果表明为鸡IL-2基因。该基因包括鸡IL-2基因的全部开放阅读框,编码142个氨基酸组成的蛋白质,与GenBank鸡IL-2基因相比,在编码氨基酸的49位有一个氨基酸缺失;而与Broiler、SC、Chenren和Xiaoshan鸡在编码氨基酸上完全一致,具有较近的亲缘关系;与Kestrel来航鸡、来航SPF鸡、Obese、Silky和Xianju鸡等有1-4个氨基酸的差异;与火鸡和鹌鹑的氨基酸同源性分别为69.9%和59.4%。将克隆到的基因插入到融合蛋白原核表达载体pGEX-6p-1中,得到重组表达质粒pGEX-IL2。将此重组质粒转化大肠杆菌DH5α,经IPTG诱导,表达出了大小约为40 ku的GST-chIL2融合蛋白,其中GST部分为26 ku,鸡IL-2为14 ku,与预期的鸡IL-2成熟蛋白大小一致。  相似文献   

4.
鸭的类似鸡白细胞介素2基因的克隆及遗传进化分析   总被引:10,自引:0,他引:10  
通过RT—PCR和DNA测序技术,克隆和测定了绍兴麻鸭的一种类似鸡白细胞介素2(IL-2)基因的核苷酸序列,并构建了该基因编码蛋白的三维结构分子模型。结果显示,绍兴麻鸭的类似于鸡IL-2基因的核苷酸长度由748个碱基组成,编码产物为由140个氨基酸残基组成的多肽,编码产物中含有21个氨基酸残基组成的信号肽。绍兴麻鸭的类似于鸡IL-2基因的核苷酸及编码产物,与鸡和火鸡IL-2的一致性分别为71.4%~72.1%和55.0%~57.9%,与哺乳类等动物IL-2的一致性分别为22.1%~28.8%和14.3%~17.1%。对绍兴麻鸭的类似于鸡IL-2基因蛋白编码产物的系统进化树分析表明,其编码产物与鸡和火鸡IL-2具有较远的亲缘关系。绍兴麻鸭的类似于鸡IL-2基因编码产物的三维结构,预测由4个α-螺旋结构和2个β折叠组成。这些研究结果预示。鸭的类似于鸡IL-2基因编码产物,在生物学功能上与哺乳动物和其他禽类具有很大的差异。  相似文献   

5.
利用生物信息学分析方法对鸡Lpin1基因编码区进行扫描,选取该基因3个引起错义突变的变异位点进行分析,其中c.391GA位点引起编码氨基酸Asp131→Asn131突变,c.1727CT位点引起编码氨基酸Ala576→Val576突变,c.2287AC位点引起编码氨基酸Met763→Leu763突变。应用引入限制性酶切位点PCR(Amplifi-cation-Created Restriction Site PCR,ACRS-PCR)的方法进行引物设计扩增3个突变位点,并通过限制性片段长度多态性(Restriction Fragment Length Polymorphism,RFLP)检测PCR产物,以藏鸡、隐性白羽鸡、藏隐(藏鸡♂×隐性白羽♀)和藏藏隐[藏鸡♂×(藏鸡♂×隐性白羽♀)♀]群体为材料对3个变异位点进行基因型检测,并与胴体性状进行关联分析;同时在5个中国地方鸡种(固始鸡、萧山鸡、北京油鸡、泰和丝羽乌骨鸡和茶花鸡)中进行变异位点的遗传多样性分析。结果表明:(1)鸡Lpin1基因c.1727CT在不同品种中具丰富的多态性;而另2个位点在所检测的群体中无多态性存在,c.391GA位点以GG基因型为主,c.2287AC位点全部为AA基因型;(2)对c.1727CT位点的遗传多样性分析表明该位点C等位基因占优势地位,且固始鸡和北京油鸡群体基因型分布显著偏离哈代-温伯格平衡定律(P0.01),在萧山鸡、泰和丝羽乌骨鸡和茶花鸡三品种中基因型分布均符合哈代-温伯格平衡定律。(3)性状关联分析显示c.1727CT位点与皮下脂肪厚呈显著关联(P0.05),与腹脂质量呈极显著关联(P0.01);其中CC基因型个体的皮下脂肪厚显著高于TT基因型个体(P0.05),极显著高于TC基因型个体(P0.01)。TT基因型个体的腹脂质量显著高于其他两种基因型个体(P0.05)。  相似文献   

6.
鸡lmbr1基因外显子16的SNP检测和单倍型分析   总被引:1,自引:0,他引:1  
外显子16是鸡lmbr1基因最大的外显子,本研究进行了该外显子在丝羽乌骨鸡和白洛克肉鸡间的SNP检测和单倍型分析。研究表明,鸡lmbr1外显子16的PCR-SSCP基因型在两个品种间的分布存在明显差异,测序从24个个体中检测到4个变异位点,其中T32C变异在两个品种间存在明显差异,丝羽乌骨鸡均含有32T(纯合的TT或杂合的TC),白洛克肉鸡在T32C位点都为纯合的CC;单倍型分析从24个个体中检测到5种单倍型,丝羽乌骨鸡和白洛克肉鸡的单倍型存在明显的差异,hap1和hap2是丝羽乌骨鸡的特异性单倍型,hap5是白洛克肉鸡的特异性单倍型,hap3和hap4主要存在于白洛克肉鸡中,在丝羽乌骨鸡中的比例极少。  相似文献   

7.
根据腺苷琥珀酸裂解酶(Adenylosuccinate lyase,ADSL)基因外显子9的序列设计引物,用PCR—SSCP的方法对隐性白羽鸡、丝羽乌骨鸡、萧山鸡、白耳鸡、藏鸡进行了单核苷酸多态性分析,并检测到了多态性,表现为3种基因型,对两种纯合子进行直接测序,结果发现9839位碱基发生了突变,由C突变为A,将核苷酸序列翻译成氨基酸序列后,发现突变后引起ADSL基因第273位氨基酸由Pro突变为Thr,为错义突变。经独立性检验,发现基因型频率和基因频率分布与品种有关,但进一步方差分析并没有显示出该位点的多态性与胸肌的IMP含量之间存在关联。  相似文献   

8.
广西东兰乌鸡由片羽和丝羽两个品系组成。目前丝羽形成的分子机理还不清楚。以前的研究表明,丝羽乌骨鸡的丝羽是由PDSS2基因上游C103G的突变所导致。本研究的目的是探讨C103G是否也是广西东兰乌鸡丝羽形成的分子基础。取广西东兰乌鸡丝羽和个体和片羽个体各20个血样,常规酚仿法提取基因组DNA,然后PCR扩增含-103位点的PDSS2片段。结果发现,广西东兰乌鸡丝羽鸡20个样的-103位点全部为G碱基,而广西东兰乌鸡片羽鸡20个样的-103位点全部为C碱基。本研究的结果提示广西东兰乌鸡丝羽形成的分子机制与丝羽乌骨鸡的相一致。  相似文献   

9.
和田黑鸡mtDNA控制区D-Loop序列分析   总被引:2,自引:2,他引:0  
试验旨在测定和田黑鸡mtDNA D-Loop序列,并分析其与其他5个品种鸡间的同源性及亲缘关系。结果发现,和田黑鸡mtDNA D-Loop序列全长为1230 bp,用DNAMAN软件进行序列比对,发现和田黑鸡与GenBank登录的5个品种鸡的同源性都在98%以上,测定样品mtDNA D-Loop核苷酸序列中富含A+T。聚类分析结果显示,和田黑鸡与丝羽乌骨鸡、红原鸡亲缘关系较近。  相似文献   

10.
用设计的特异性引物,采用RT-PCR技术特异性地扩增到丝羽乌骨鸡β-防御素基因Gal-1(Gallinacin-1),Gal-1(a)(Gallinacin-1(a)),Gal-2(Gallinacin-2),Gal-4(Gallinacin-4)~Gal-13(Gallinacin-13)基因共13个基因编码区全长片段。通过克隆、测序获得13个基因的cDNA核苷酸序列,并提交到GenBank,丝羽乌骨鸡β-防御素Gal-1,Gal-1(a),Gal-2,Gal-4~Gal-13基因的登录号为:DQ677632 ̄DQ677644。将丝羽乌骨鸡13种β-防御素Gal-1~Gal-13基因分别与GenBank中收录的防御素基因比对分析,其氨基酸序列的相似性均在95.5%~100%之间。利用DNAStar软件对所获得的13种丝羽乌骨鸡β-防御素基因进行系统发育树分析,结果Gal-1、5、12在同一分支上,Gal-1(a)、4、8在同一分支上,Gal-1、2、9在同一分支,Gal-6、7在同一分支,Gal-13独在一分支上。  相似文献   

11.
丝羽乌骨鸡与其它鸡种遗传关系的研究   总被引:9,自引:0,他引:9  
乌骨鸡是著名的药用品种 ,本文通过测定乌骨鸡和其它三种家鸡的线粒体DNAD -环区部分序列来研究其遗传结构与其它鸡种的异同。在所测的 2 0个DNA序列中共检测到 2 4个变异位点 ,变异率为 4.45 % ,说明乌骨鸡与其它鸡种的差异较小。研究还显示 ,乌骨鸡与白银耳鸡关系最近、与萧山鸡关系次之 ,与仙居鸡距离较远  相似文献   

12.
Interleukin (IL)-1beta-encoding regions of chicken, duck, goose, turkey and pigeon were cloned and sequenced. Each IL-1beta-encoding region of chicken, duck, goose and turkey is 804 nucleotides long and encodes IL-1beta protein that is 268 amino acids. Pigeon IL-1beta-encoding region is 810 nucleotides long and encodes IL-1beta protein that is 270 amino acids. Two one-nucleotide and one four-nucleotide insertions of pigeon IL-1beta-encoding region sequence were found, resulting in two amino acid insertions in pigeon IL-1beta. Pairwise sequence analysis showed that the sequence identities of IL-1beta-encoding genes ranged from 77% to 99%, which were also found for IL-1beta protein sequence identities, with an average level of both sequence identities of 89%. Phylogenetic analysis indicated that IL-1beta-encoding regions and the encoded proteins of chicken, duck, goose and turkey clustered together and evolved into a distinct phylogenetic lineage from that of pigeon which evolved into a second lineage. The results from the binding reaction of antiserum against each recombinant IL-1beta (r IL-1beta) protein to homologous or heterologous rIL-1beta, the enhancement levels of K60 mRNA expression in rIL-1beta-treated DF-1 cells or the reduction levels of K60 mRNA expression in DF-1 cells treated with rIL-1beta that was preincubated with homologous or heterologous antiserum showed that all five rIL-1beta were functional active and shared significantly structural and functional homology.  相似文献   

13.
根据GenBank上已发表的犬科动物的白细胞介素-2(IL-2)基因序列,在开放阅读框架上下游的保守区域设计了1对引物。无菌采取貉血,使用淋巴细胞分离液分离外周血液中的淋巴细胞,加入ConA,于二氧化碳培养箱中培养48h后收集培养细胞。以培养细胞中提取的总RNA为模板,应用RT—PCR方法,扩增出貉的IL-2基因。分离纯化片段,连接T载体转化大肠杆菌并测序。结果表明:IL-2基因开放阅读框架全长486bp,编码155个氨基酸。该序列与大、狐等犬科动物的IL-2基因同源关系最近,与大熊猫、家猫的IL-2基因同源关系相对较近,与人、马、牛、绵羊的IL-2基因有一定的遗传距离,与禽类鸡的IL-2基因同源关系最远。  相似文献   

14.
杂交猪白细胞介素-18全基因的克隆与序列分析   总被引:1,自引:0,他引:1  
通过RT-PCR方法直接从猪脾脏淋巴细胞中扩增出猪白细胞介素-18(IL-18)全基因的cDNA,其大小为579bp,编码192个氨基酸。与GenBank上已发表猪IL-18序列(ABO10003)进行比较,核苷酸同源性为99.8%,在第550位处(以ATG为1计)由A→G,存在有意义突变。与GenBank上的ABO10003、AF176949、AY262109、NM1997序列进行比较分析,氨基酸同源性分别为99%,98.5%,99.8%和99%。从系统进化树可以看出,河南良杂猪IL-18基因与ABO10003、AY262109亲缘关系最近。河南良杂猪IL-18基因和人及其他动物IL-18基因核苷酸序列进行比较分析,结果显示猪与人、猫、牛、鸡、鸭、海豚、山羊、马、家鼠、老鼠、绵羊的IL-18基因核苷酸同源性分别为83.1%、88.3%、90.7%、26.8%、31.4%、26.3%、90.0%、91.5%、67.7%、65.1%和90.8%。  相似文献   

15.
The study was mainly focused on the genetic characteristics of frizzled feather trait,the phenotype difference between frizzled feather(FF)and incomplete frizzled feather(IFF),and SNP analysis of the candidate gene KRT75 in the frizzle and incomplete frizzle chickens of different feather colors,which could provide scientific gist for the frizzle gene mapping,development and utilization of the two kinds of chicken.The resource population(homozygous Yellow feather Kirin chicken×Huaixiang chicken)of hybrid F2 was established.The back,neck,wing primary and wing-bow feather were observed and the scatter diagram of rachis bending degree was drawn.The blood of 10 white frizzle feather Kirin(WFF)chicken,10 Yellow frizzle feather Kirin(YFF)chicken,10 Royal,25 Yellow feather incomplete frizzle(YFIF)chicken and 15 Black feather incomplete frizzle(BFIF)chicken from homozygous WFF chicken×Royal chicken of hybrid F1 were extracted,then the DNA of those samples were extracted.The primer of KRT75 was designed,the PCR products were also sequenced after purification,Chromas 2.22 and DNAMAN software were used to analysis of sequence peaks photos and sequence alignment,respectively.The amount of orthogonal and reciprocals cross in hybrid F1 was 127 and 139, respectively, which all were the slight(incomplete)frizzled feather phenotype,with no different penetrance.The resource population of hybrid F2 were consisted of 55 frizzle chicks including some hens and cocks,106 incomplete frizzled and 68 contour feather chicken,the proportion of which was coincided with Mendelian segregation ratio of 1:2:1 by χ2-test(P>0.05),therefore it preliminary verified that the frizzle gene F was autosomal incomplete dominant inheritance.FF and IFF curved upward deviating from the skin,and the line slope of trend line of FF was 3.5 times than that of IFF.It found that there was no 69 bp deletion mutation in exon 5 region of KRT75 gene in WFF and YFF chicken,but 3 SNPs(T71C,T83C,C95T)were deleted in the 69 bp,which were homologous CC in WFF and YFF chicken,of two were heterozygous in YFIF chicken,while all were heterozygous in BFIF chicken.2 SNPs(T662C and T770C)were also deleted in exon 6,heterozygous of which were only deleted in BFIF chicken.The haplotype analysis indicated that 9 haplotypes were detected in 60 individuals,hap1/hap1 was specific genotype of WFF and YFF chicken.The haplotypes of two incomplete frizzle feather chicken were apparently different,hap4 was specific haplotype of YFIF chicken,while hap6,hap7,hap8 and hap9 were specific haplotypes of BFIF chicken.The frizzle gene F was autosomal incomplete dominant inheritance,there were obvious differences between FF and IFF traits,5 SNPs in exon 5 and 6 of KRT75 gene probably were the reference of molecular markers as distinguishing between frizzle and incomplete frizzle chicken.  相似文献   

16.
Silky fowl, a breed of chicken, is hyperpigmented in its various internal tissues. The pigment was extracted from various tissues of two strains of Silky fowl to determine its molecular structure and internal distribution. Analysis by infrared spectroscopy showed two spectrum patterns of the pigment in Silky fowl; one is from ovary and testis, the other is from periosteum and feather. The difference between the two spectra is possibly due to the sulfur contents of melanin. Especially, the spectra of the pigments from feather and periosteum shared the characteristics of synthesized melanin spectrum in common, which indicates that the melanocytes dispersed in these tissues were functionally the same. According to our quantitative analysis, the tissues examined were classified significantly in the order of the pigment content (p<0.05): periosteum > gonads (ovary or testis) = trachea > or = heart, liver, gizzard, cecum, muscles (Pectoralis and Supracoracoideus) and skin. In addition, the specific regions of embryonic neural crest derived cells, such as cardiac artery and various parts of cephalic tissues, were found to be locally hyperpigmented. These data suggest that hyperpigmentation (fibromelanosis) in Silky fowl chicken occurs in a tissue- and organ-specific manner, which is strongly related to neural crest cell development. It is hypothesized that neural crest cells of the bird, containing melanocyte progenitors, acquire unusual ability to differentiate into melanocytes excessively, and to extend the distribution of their descendant along the destinations of neural crest derivatives.  相似文献   

17.
利用7个微卫星标记对12个贵州地方鸡种及1个引入鸡种的遗传多样性进行了检测,分析了各标记座位上的等位基因及频率、基因杂合度、平均基因杂合度、多态信息含量、平均多态信息含量及群体间的亲缘关系。结果表明:13个种群在7个标记座位上的等位基因及频率等均存在一定差异。其中,瑶山鸡的平均基因杂合度最高,引入鸡种隐性白洛克的平均基因杂合度最低,其余鸡种介于其间。平均多态信息含量的分析结果与此类似,说明瑶山鸡的遗传多样性最为丰富。模糊聚类的结果表明,13个鸡种分为2大类群,隐性白洛克独自为1个类群,12个贵州地方鸡种构成1个类群。12个贵州地方鸡种又可进一步分为2个小类群:其中1个小类群由兴义土鸡、矮脚鸡首先聚在一起,然后依次聚上黔东南小香鸡、黔东南小香乌骨鸡构成;高脚鸡、瑶山鸡、威宁鸡、竹乡鸡、竹乡乌骨鸡、黑羽乌蒙乌骨鸡、乌蒙鸡、麻羽乌蒙乌骨鸡则构成另一个小类群。  相似文献   

18.
利用27个微卫星DNA标记对12个地方乌骨鸡品种进行遗传多样性分析,通过计算等位基因频率、多态信息含量、杂合度、有效等位基因数和遗传距离,评估各品种内遗传变异和各品种间遗传相关,并根据遗传距离对12个鸡品种进行了聚类分析。结果表明,各乌骨鸡群体的遗传多样性较为丰富,并具有较高的选择潜力。平均杂合度最高的是竹乡鸡,为0.670;最低的是江山乌骨鸡,为0.581。27个微卫星标记中4个为中度多态座位,1个低度多态座位,22个为高度多态座位;LEI0234与LEI0192分别检测到了10.3与12.2个等位基因。以标准遗传距离为准,遗传距离最近的为沐川乌骨鸡与略阳鸡,为0.1002,而乌蒙乌骨鸡与略阳鸡最远,为0.2546。通过UPG—MA法聚类后,略阳鸡、沐川乌骨鸡、兴文乌骨鸡、盐津乌骨鸡、竹乡鸡首先聚为1类,江山乌骨鸡、郧阳白羽乌鸡、余干乌骨鸡聚为1类;金湖乌凤鸡、丝绒乌骨鸡和丝毛乌骨鸡聚为第3类;乌蒙乌骨鸡独为一类。  相似文献   

19.
试验旨在探究多个地方品种快慢羽鸡的内源性病毒基因21(ev21)以及SPEF2基因与PRLR基因的部分重复序列(JS序列)分布对羽速基因(Kk)表型的影响。采用PCR扩增、HaeⅢ限制性内切酶酶切的方法检测不同地方品种快慢羽鸡性染色体上OR区域(ev21占据片段)、UR区域(URa、URb(ev21未占据片段))以及JS序列分布情况。结果表明:国内的一些地方品种部分慢羽鸡个体OR区域ev21基因缺失以及部分快羽鸡的URb区域存在ev21基因插入;可通过JS序列扩增对快慢羽表型进行准确鉴定。研究结果显示JS序列可作为快慢羽鉴定候选基因,该方法可广泛应用于国内快慢羽鸡种的鉴定。  相似文献   

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