鸡减蛋综合症病毒（EDSV）基因组右末端结构的分析

从中国发病鸡中分离的鸡减蛋综合征病毒（ＥｇｇＤｒｏｐＳｙｎｄｒｏｍＶｉｒｕｓ,ＥＤＳＶ）弱毒株（ＡＡ－２）,其基因组全长约为３３ｋｂ。用限制性内切酶ＨｉｎｄⅢ水解ＥＤＳＶ全基因组,构建了以ｐＢｌｕｅｓｃｒｉｐｔⅡ（ＫＳ＋）为载体的右末端片段的克隆。对其进行了序列 测定和结构分析。     

藏鸡线粒体全基因组序列的测定和分析

通过PCR扩增,测序,拼接,获得藏鸡（Tibetan Chicken）线粒体全基因组序列并进行数据分析处理。藏鸡线粒体全基因组序列全长16783bp,共有13个蛋白质编码基因、2个rRNA基因、22个tRNA基因和1个D-loop区。模拟电子酶切结果显示,藏鸡DraI酶的酶切结果和先前报道的原鸡,茶花鸡,尼西鸡和大理漾濞黄鸡的酶切结果都不相同,为藏鸡特有。基于D-loop区全序列和13个蛋白质编码基因序列,采用N-J算法与原鸡属4个种,3个亚种和3个家鸡品系构建系统进化树：初步确定藏鸡起源于红原鸡,与家鸡中的来航鸡、白洛克鸡亲缘关系最近,但是藏鸡的进化与来航鸡、白洛克鸡这两个家鸡品系又显得相对独立。推测可能原因是藏鸡的祖先在进入高原以后处于相对封闭的环境,从而形成了独特群体遗传特性。     

麦迪霉素4″—O—丙酰基转移酶（mpt）基因结构的研究

对含有麦迪霉素４″－Ｏ－丙酰基转移酶（ｍｐｔ）基因的ＢａｍＨＩ－ＢａｍＨＩ８．０ｋｂ的ＤＮＡ片段进行限制性酶切分析,绘制出了含有２１个酶切位点的限制性酶切图谱。以含有碳霉素异戊酰基转移酶基因（ＣａｒＥ）的２．４ｋｂ ＤＮＡ片段为探针,经Ｓｏｕｔｈｅｒｎ ｂｌｏｔ分子杂交,将ｍｐｔ定位于一个ＥｃｏＲＩ－ＥｃｏＲＩ－ＰｓｔＩ３．０ｋｂ的ＤＮＡ片段上,将该片段克隆至大肠杆菌／链霉菌穿梭质粒载体ｐＷＨＭ３     

隼形目鹰科四种鸟类线粒体DNA研究

本实验采用ＡｐａＩ,ｆＢａｍＨＩ,ＢｇｉＩＩ,ＥｃｏＲＩ,ＥｃｏＲＶ,ＨｉｎｄⅢＨｐａＩ,ＫｐｎＩ,ＰｓｔＩ,ＰｒｕＩＩ,ＳａｌＩ,ＳｃａＩ,ＸｂａＩ和ＸｈｏＩ１４种限制性内切酶,对鹰科４种鸟类即雀鹰（Ａｃｃｉｐｉｔｅｒｎｉｓｕｓ）,松雀鹰（Ａ．ｖｉｒｇａｔｕｓ）苍鹰（Ａ．ｇｅｎｔｉｌｉｓ）和灰险鹰（Ｂｕｆｔａｔｕｒｉｎｄｉｃｕｓ）ｍｔＤＮＡ限制性片段长度多态分析。结果表明,４种鸟类中雀鹰和松１主     

鸡减蛋综合征病毒(EDSV)基因组右末端结构的分析

从中国发病鸡中分离的鸡减蛋综合征病毒（ＥｇｇＤｒｏｐＳｙｎｄｒｏｍＶｉｒｕｓ,ＥＤＳＶ）弱毒株（ＡＡ－２）,其基因组全长约为３３ｋｂ。用限制性内切酶ＨｉｎｄⅢ水解ＥＤＳＶ全基因组,构建了以ｐＢｌｕｅｓｃｒｉｐｔⅡ（ＫＳ＋）为载体的右末端片段的克隆（约４．２ｋｂ）,对其进行了序列测定和结构分析。该片段全长４１８３个碱基对（ｂｐ）,位于基因组右末端８７．３ｍ．ｕ．－１００ｍ．ｕ．。结果显示,该片段与哺乳动物腺病毒右末端Ｅ４区结构不同,与禽Ⅰ型腺病毒代表株ＣＥＬＯ右末端片段亦无同源性。本文为深入了解ＥＤＳＶ基因组结构特点,ＥＤＳＶ与其他腺病毒基因结构与功能的进化关系和ＥＤＳＶ载体的构建奠定了分子生物学基础     

板齿鼠线粒体DNA的研究

本文应用ＡｐａＩ、ＢａｍＨＩ、ＢｃｌＩ、ＢｇｌＩ、ＣｌａＩ、ＥｃｏＲＩ、ＥｃｏＲＶ、ＨｉｎｄＩＩ、ＰｓｔＩ、ＰｖｕＩＩ、ＳａｃＩ、ＳｃａＩ和ＸｂａＩ等１３种限制性内切酶对板齿鼠线粒体ＤＮＡ（ｍｔＤＮＡ）进行限制性片段长度多态（ＲＦＬＰ）分析,并用双酶解法构建其限制性内切酶图谱。结果表明板齿鼠存在３种ｍｔＤＮＡ单倍型,可通过限制酶ＰｖｕＩＩ、ＨｉｎｄＩＩ和ＡｐａＩ区分,呈现ＤＮＡ多态性和种内遗传变异。与小家鼠、褐家鼠ｍｔＤＮＡ限制性片段的数据相比较,板齿鼠和这两种鼠ｍｔＤＮＡ存在明显差异。板齿鼠ｍｔＤＮＡ限制性内切酶图谱的建立,为进一步系统研究鼠科动物的遗传分化提供了依据。     

日本血吸虫26kD抗原基因在BCG中的表达

研究了外源基因日本血吸虫２６ｋＤ抗原（Ｓｊ２６ＧＳＴ）在卡介苗（ｂａｃｉｌｕｓＣａｌｍｅｔｅ－Ｇｕｅｒｉｎ,ＢＣＧ）、耻垢分枝杆菌（Ｍ．ｓｍｅｇｍａｔｉｓ）和大肠杆菌（Ｅ．ｃｏｌｉ）中的表达．运用重组ＤＮＡ和聚合酶链反应（ＰＣＲ）等分子生物学技术,以表达Ｓｊ２６ＧＳＴ的Ｅ．ｃｏｌｉｐＧＥＸ衍生质粒为模板,经ＰＣＲ得到编码Ｓｊ２６ＧＳＴ的全长ｃＤＮＡ片段．将其按正确的阅读框顺序,克隆到人结核杆菌热休克蛋白（ｈｅａｔｓｈｏｃｋｐｒｏｔｅｉｎ,ＨＳＰ）７０的启动子下游,再将ＨＳＰ７０启动子和Ｓｊ２６ＧＳＴ基因一起亚克隆到Ｅ．ｃｏｌｉ－分枝杆菌穿梭质粒ｐＢＣＧ－２０００中,得到Ｅ．ｃｏｌｉ－分枝杆菌穿梭表达质粒ｐＢＣＧ－Ｓｊ２６．ｐＢＣＧ－Ｓｊ２６电转化入ＢＣＧ和Ｍ．ｓｍｅｇｍａｔｉｓｍｃ２１５５中表达Ｓｊ２６ＧＳＴ抗原,所表达的天然重组Ｓｊ２６ＧＳＴ（ｒＳｊ２６ＧＳＴ）为可溶性蛋白,在ＳＤＳ－ＰＡＧＥ上分子量为２６ｋＤ处可见明显的表达蛋白带．其表达量分别占ＢＣＧ和Ｍ．ｓｍｅｇｍａｔｉｓ菌体总蛋白的１５％和１０％．可见,Ｓｊ２６ＧＳＴ基因能在ＢＣＧ中高效表达．     

自发性高血压大鼠原癌基因表达和限制性内切酶片段长度多态性分析

观察自发性高血压大鼠（ＳＨＲ）血管平滑肌细胞（ＶＳＭＣｓ）的生长曲线、ｃ－ｆｏｓ原癌基因表达和ｃ－ｆｏｓ基因酶切图谱的情况,与对照组京都维斯特大鼠（ＷＫＹ）进行对比。结果显示,在小牛血清作用下ＳＨＲ的ＶＳＭＣｓ生长速率和ｃ－ｆｏｓ原癌基因表达明显大于ＷＫＹ;ｃ－ｆｏｓ原癌基因限制性内切酶片段长度多态性（ＲＦＬＰ）分析表明,经ＢａｍＨ Ｉ或ＥｃｏＲ Ｉ酶切后的ＳＨＲ－ＷＫＹ的酶切图谱一致,同时也未发     

树鼠mtDNA限制性内切酶图谱及其与小家鼠褐家鼠的亲缘关系

本实验用ＡｐｅⅠ,ＡｖａⅠ,ＢａｍＨⅠ,ＢｃｌⅠ,ＢｇｌⅠ,ＣｌａⅠ,ＥｃｏＲⅠ,ＥｃｏＲＶ,ＨｐａⅠ,ＰｓｔⅠ,ＰｖｕⅡ,ＳｃａⅠ,ＸｂａⅠ等１３种限制性内切酶分析树鼠（Ｃｈｉｒｏｍｙｓｃｕｓｃｈｉｒｏｐｅｓ）的ｍｔＤＮＡ限制性片段长度多态性,并用双酶解法构建了其中８种酶的限制性内切酶图谱。根据限制性片段差异法和分子钟,计算并讨论树鼠和小家鼠（Ｍｕｓｍｕｓｃｕｌｕｓ）、褐家鼠（Ｒａｔｔｕｓｎｏｒｖｅｇｉｃｕｓ）的ｍｔＤＮＡ遗传距离和亲缘关系。结果表明树鼠与褐家鼠的关系较接近,两者的分歧时间在距今１５００─２０００万年前,即处于中新世早中期。     

贵州汉族,苗族,布依族和水族人群线粒体DNA多态性研究

本文运用１６种限制性内切酶对来自贵州的汉族、苗族、布依族和水族的１５０个样本进行了ｍｔＤＮＡ的限制性片段长度多态性（ＲＦＬＰ）分析。共检测到３１种限制性格局（Ｒｅｓｔｒｉｃｔｉｏｎｐａｔｔｅｒｎ）,其中ＨａｅＩＩ－１３型、ＥｃｏＲＶ－３型和ＰｓｔＩ－４型３种限制性格局为新报道综合这些限制性格局,共得出２８种ｍｔＤＮＡ类型（ｍｔＤＮＡｔｙｐｅ）。运用ＵＰＧ法和简约法分析了各ｍｔＤＮＡ类型之间、各人群之间的聚类关系,结果表明：水族人群的ｍｔＤＮＡ变异度较大;汉族和苗族的亲缘关系最近,布依族和水族有着较远的亲缘关系。     

Analysis of genetic diversity and phylogenetic relationships among red jungle fowls and Chinese domestic fowls

Genetic diversity and phylogenetic relationships among 568 individuals of two red jungle fowl subspe- cies (Gallus gallus spadiceus in China and Gallus gallus gallus in Thailand) and 14 Chinese domestic chicken breeds were evaluated with 29 microstaellite loci, the genetic variability within population and genetic differentiation among population were estimated, and then genetic diversity and phylogenetic relationships were analyzed among red jungle fowls and Chinese domestic fowls. A total of 286 alleles were detected in 16 population with 29 microsatellite markers and the average number of the alleles observed in 29 microsatellite loci was 9.86±6.36. The overall expected heterozygosity of all population was 0.6708±0.0251, and the number of population deviated from Hardy-Weinberg equilibrium per locus ranged from 0 to 7. In the whole population, the average of genetic differentiation among population, measured as FST value, was 16.7% (P<0.001), and all loci contributed significantly (P<0.001) to this differentiation. It can also be seen that the deficit of heterozygotes was very high (0.015) (P<0.01). Reynolds' distance values varied between 0.036 (Xiaoshan chicken-Luyuan chicken pair) and 0.330 (G. gallus gallus-Gushi chicken pair). The Nm value ranged from 0.533 (between G. gallus gallus and Gushi chicken) to 5.833 (between Xiaoshan chicken and Luyuan chicken). An unrooted consensus tree was constructed using the neighbour-joining method and the Reynolds' genetic distance. The heavy-body sized chicken breeds, Luyuan chicken, Xiaoshan chicken, Beijing Fatty chicken, Henan Game chicken, Huainan Partridge and Langshan chicken formed one branch, and it had a close genetic relationship between Xiaoshan chicken-Luyuan chicken pair and Chahua chicken-Tibetan chicken pair. Chahua chicken and Tibetan chicken had closer genetic relationship with these two subspecies of red jungle fowl than other domestic chicken breeds. G. gallus spadiceus showed closer phylogenetic relationship with Chinese domestic chicken breeds than G. gallus gallus. All 29 microstaellite loci in this study showed high levels of polymorphism and significant genetic differentiation was observed among two subspecies of red jungle fowl and 14 Chinese domestic chicken breeds. The evolutional dendrogram is as follows: evolutional breeds→primitive breeds (Chahua chicken and Tibetan)→red jungle fowl in China (G. gallus spadiceus)→red jungle fowl in Thailand (G. gallus gallus). The results supported the theory that the domestic fowls might originate from different subspecies of red jungle fowl and Chinese domestic fowls had independent origin.     

家鸡腺苷琥珀酸裂解酶基因多态性与肌苷酸含量的关联及其与红原鸡的亲缘关系

根据腺苷琥珀酸裂解酶(adenylosuccinate lyase, ADSL)基因外显子2的序列设计引物, 用PCR-SSCP的方法对隐性白羽鸡、丝羽乌骨鸡、白耳鸡、藏鸡以及红色原鸡两个亚种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3484位碱基处发生C→T突变。对3种基因型的肌肉肌苷酸含量的最小二乘分析结果显示TT型(突变型)个体的肌肉肌苷酸含量极显著地高于CT型、显著地高于CC型个体, CT型个体也稍高于CC型, 但差异不显著, 初步推测该位点可能与肌肉肌苷酸含量有关。根据该多态位点的基因频率, 基于Nei氏的遗传距离运用NJ聚类法构建系统发生树, 进行家鸡与原鸡的亲缘关系分析, 结果发现, 丝羽乌骨鸡与白耳鸡的亲缘关系最近, 藏鸡和中国红原鸡亚种的亲缘关系也较近, 中国地方家鸡品种与中国红原鸡亚种的亲缘关系较近,而与泰国红色原鸡的亲缘关系较远,隐性白羽鸡与原鸡亲缘关系最远, 初步得出中国家鸡有自己独自的血缘来源的结论。     

A variant in the restriction endonuclease cleavage pattern of mitochondrial DNA in the domestic fowl, Gallus gallus domesticus

Summary. The cleavage patterns of mitochondrial DNAs (mtDNAs) were investigated from 15 lines of domestic fowls, Gallus gallus domesticus . using 11 restriction endonucleases. The cleavage patterns with 10 restriction endonucleases were identical in all the lines. A variant was found in a line of White Leghorn in the pattern with Mspl digestions. Cleavage patterns of the red jungle fowl, Gallus gallus gallus , were identical to the common patterns shown by the 14 lines of domestic fowls.     

中国红原鸡和泰国红原鸡遗传多样性分析

利用29个微卫星DNA标记对来自中国的红原鸡Gallus gallus spadiceus亚种和来自泰国的红原鸡Gallus gallus gallus亚种进行遗传多样性分析, 评估亚种内的遗传变异和亚种间的遗传分化, 结果表明: 共检测到168个等位基因, 每个位点的等位基因数从2到13不等, 所有位点平均的期望杂合度和PIC值分别为0.5780和0.53。中国和泰国红原鸡29个微卫星位点平均有效等位基因数分别为3.79和4.79, 平均基因杂合度为0.5379和0.6385, 两个红原鸡亚种均表现出较高的群体杂合度和丰富的遗传多样性。群体分化系数为19.4%(P<0.01), 两个红原鸡亚种间的Reynolds’遗传距离和Nm值分别为0.157和1.040。由此可见, Gallus gallus spadiceus亚种和Gallus gallus gallus亚种群体具有不同的群体遗传结构, 群体之间存在明显的遗传分化, 并不能将其认定为是同一亚种, 这也为中国家鸡具有独立的起源提供了一定的佐证。     

Restriction Enzyme Analysis of Mitochondrial DNA of Acanthamoeba Strains in Japan

ABSTRACT. Eight isolates, identified as either Acanthamoeba castellanii or A. polyphaga from human eye infections, contact lens containers, and soil in Japan, were characterized by restriction fragment length polymorphisms (RFLP) of mitochondrial DNA (mtDNA). Mitochondrial DNA was digested with either Bgl II, Eco R I, Hind III, Hpa I, Sca I or Xba I, electrophoresed in agarose gels, and stained with ethidium bromide. Four distinct RFLP phenotypes that refer to the collection of six fragment size patterns obtained for a single strain with six enzymes, were discovered among the eight strains used in this study. Three strains morphologically classified as A. polyphaga share a single RFLP phenotype with the Ma strain of A. castellanii. The interspecific sequence differences of 7.06–12.74% in DNA nucleotide were estimated from the proportion of DNA fragments shared by each pair of mtDNA.     

Restriction endonuclease analysis of mitochondrial DNA of three farm animal species: Cattle, sheep and goat

Five restriction endonucleases (HindIII, BgIII, EcoRI, EcoRV and BamHI) were employed to analyse mitochondrial DNA of cattle, sheep and goat. The results showed completely different restriction patterns of mtDNA among the three bovid species. A total of 11, 16, and 17 restriction fragments in cattle, sheep and goat respectively, were detected by the five restriction endonucleases. Average total sizes of mtDNA of cattle, sheep and goat were found to be 16.49 ± 0.18, 16.30 ± 0.25 and 16.44 ± 0.08 kb, respectively. The mtDNA cleavage patterns were identical for all seven individuals belonging to two cattle breeds and for 10 individuals from one sheep breed.     

海南文昌鸡若干种质特征和遗传多样性研究

对产于热带亚热带的文昌鸡若干种质特性进行了测定,采用RAPD技术分析其遗传多样性,并与其它生态类型鸡种进行了比较研究．结果表明,文昌母鸡羽毛以棕褐色、黄褐色和黄麻色为主,占群体80．83％;公鸡以红色、背翼部夹杂黑羽为主,占群体71．96％．文昌鸡的胚色大多为黄色．文昌鸡的生长速度比其它生态类型鸡种较慢,40周龄公母鸡的体重、体斜长和胚长也较小,明显具有热带亚热带生态特征．文昌鸡性早熟,母鸡127d产蛋率5％,182d产蛋率50％,300d产蛋量73个,平均蛋重37．6±3．2g．对文昌鸡基因组DNA进行RAPD分析,鸡群个体或群体间平均相似系数为0．6071-0．9699,表明文昌鸡的同源性普通高于内地其它鸡种,保持了品种的纯度．     

Insights into the genetic history of Green‐legged Partridgelike fowl: mtDNA and genome‐wide SNP analysis

The Green‐legged Partridgelike (GP) fowl, an old native Polish breed, is characterised by reseda green‐coloured shanks rather than yellow, white, slate or black commonly observed across most domestic breeds of chicken. Here, we investigate the origin, genetic relationships and structure of the GP fowl using mtDNA D‐loop sequencing and genome‐wide SNP analysis. Genome‐wide association analysis between breeds enables us to verify the genetic control of the reseda green shank phenotype, a defining trait for the breed. Two mtDNA D‐loop haplogroups and three autosomal genetic backgrounds are revealed. Significant associations of SNPs on chromosomes GGA24 and GGAZ indicate that the reseda green leg phenotype is associated with recessive alleles linked to the W and Id loci. Our results provide new insights into the genetic history of European chicken, indicating an admixd origin of East European traditional breeds of chicken on the continent, as supported by the presence of the reseda green phenotype and the knowledge that the GP fowl as a breed was developed before the advent of commercial stocks.     

Restriction cleavage map of mitochonrial DNA from the yeast Saccharomyces cerevisiae.

Mitochondrial DNA (mtDNA) from the yeast Saccharomyces cerevisiae was cleaved by restriction endonucleases Eco RI, Hpa I, Bam HI, Hind III, Pst I, and Sal I, yielding 10, 7, 5, 6, 1, and 1 fragments, respectively. A physical ordering of the restriction sites on yeast mtDNA has been derived. Yeast mtDNA cannot be isolated as intact molecules, and it contains nicks and gaps which complicate the use of conventional fragment mapping procedures. Nevertheless, the position of each of the restriction sites was obtained primarily by reciprocal redigestion of isolated restriction fragments. This procedure was supplemented by co-digestion of mtDNA with a multisite enzyme and a single-site enzyme (i.e., Sal I or Pst I) which provided a unique orientation for overlapping fragments cleaved by Sal I or Pst I. The data obtained from these approaches were confirmed by analysis of double and triple enzyme digests. Analysis of partial digest fragments was used for positioning of the smallest Eco RI fragment. A comparison of mtDNA from four grande strains (MH41-7B, 19d, TR3-15A, and MH32-12D) revealed similar, but slightly varying restriction patterns, with an identical genome size for each of approximately 5 X 10(-7) d or 75 kb. A fifth grande strain, D273-10B from S. cerevisiae, revealed restriction patterns different from those of the above strains, with a smaller genome size of 70 kb.     

Domestic chickens defy Rensch's rule: sexual size dimorphism in chicken breeds

Sexual size dimorphism (SSD), i.e. the difference in sizes of males and females, is a key evolutionary feature that is related to ecology, behaviour and life histories of organisms. Although the basic patterns of SSD are well documented for several major taxa, the processes generating SSD are poorly understood. Domesticated animals offer excellent opportunities for testing predictions of functional explanations of SSD theory because domestic stocks were often selected by humans for particular desirable traits. Here, we analyse SSD in 139 breeds of domestic chickens Gallus gallus domesticus and compare them to their wild relatives (pheasants, partridges and grouse; Phasianidae, 53 species). SSD was male-biased in all chicken breeds, because males were 21.5 ± 0.55% (mean ± SE) heavier than females. The extent of SSD did not differ among breed categories (cock fighting, ornamental and breeds selected for egg and meat production). SSD of chicken breeds was not different from wild pheasants and allies (23.5 ± 3.43%), although the wild ancestor of chickens, the red jungle fowl G. gallus, had more extreme SSD (male 68.8% heavier) than any domesticated breed. Male mass and female mass exhibited positive allometry among pheasants and allies, consistently with the Rensch's rule reported from various taxa. However, body mass scaled isometrically across chicken breeds. The latter results suggest that sex-specific selection on males vs. females is necessary to generate positive allometry, i.e. the Rensch's rule, in wild populations.