五指山猪生物学特性、易地繁育及遗传多样性研究

五指山猪是我国濒临灭绝的珍稀品种,为保护和利用这一特有种猪基因资源,我们开展了易地繁育和生物学特性及遗传多样性的研究。通过对其生长发育规律、繁殖生理、血液生理生化、遗传标记、DNA指纹图谱等项的观察和测试,发现五指山猪具有体型小、性成熟早、耐近交繁殖、遗传性较稳定、肉质好等特点。白细胞分类中淋巴细胞占77％,白细胞抗原(SLA) 血清学分型中,与其它品种之间存在有差异性。 用人源小卫星探针33.6和鸡小卫星探针cMS18,对五指山猪及长白猪和枫泾猪进行DNA指纹图比较分析,发现五指山猪的DNA指纹图相似系数,大大高于其它已研究过的正常体型猪种,说明它经历过较高程度的近交;以猪的生长激素（GH）cDNA为探针,用EcoRI、EcoRV、PstI等酶切基因组DNA进行RFLP分析时,五指山猪均出现了一条深色带,而长白猪和枫泾猪未发现。表明五指山猪在生长激素位点上与正常体型猪种亦存在差异。     

枫泾猪、香猪和长白猪的DNA指纹图分析

本文用M13和3个人源小卫星探针(33.6、 33.15、α-珠蛋白-3’HVR)获得了枫泾猪、香猪及长白猪的DNA指纹图,分析了各品种内和品种间的遗传变异性。结果表明,各品种的平均DNA指纹图相似系数在0.440-0.532的范围内,极显著地高于品种间的相似系数(0.223-0.307)。各品种的DNA指纹图还存在着一些品种特异性图带,这些图带在以后的基因定位中值得重视。 Abstract:This paper demonstrated DNA fingerprints of Fengjing,Xiang and Large White breeds of swine generated by M13 and three human minisatellite probes(33.6,33.15 andα-globin-3’HVR),and determined the intrabreed and interbreed variabilities of swine DNA fingerprints.It was shown that the mean band sharing probability within breeds was 0.440-0.532,significantly higher than that between breeds(0.223-0.307).Some breed-specific bands were noted in each breed,which should draw our attention in the course of gene mapping.     

中国三种实验用小型猪线粒体DNAD-loop多态性分析

分析中国三种实验用小型猪线粒体DNA（mtDNA）D-loop的多态性,建立各品种品系猪的遗传标记,为各品种、品系猪的鉴别提供依据。应用PCR技术分别对西双版纳近交系小耳猪、广西巴马小型猪、贵州小型香猪和长白猪的血液总DNA样品中mtDNAD-loop进行扩增,用23种限制性内切酶消化,观察其酶切多态。PCR扩增其mtDNAD-loop5′端227bp高变区域,应用PCR?SSCP和PCR直接测序分析,观察其单链构象多态和序列多态。结果显示：三种小型猪之间未见酶切长度多态、单链构象多态和序列多态。与长白猪之间表现出单链构象多态和序列多态。本研究认为：三种实验用小型猪之间mtDNA多态性贫乏,证明其亲源关系很近,在母系起源和进化上有一致性,应用PCR RFLP、PCR SSCP和PCR直接测序分析,尚不能作为三种实验用小型猪品种、品系鉴定的依据,但与长白猪等欧系猪比较有一定差异。 Abstract：the present study is to analyze the polymorphism of the mtDNA D-loop in three breeds of laboratory miniature pigs in China , and to establish its cytoplasmic DNA markers to distinguish among them . The polymorphism of mtDNA D-loop and its 5′-end high variable regions were detected by PCR-RFLP , PCR-SSCP and PCR-direct sequencing on Xishuangbanna Small-ear inbred pig, Guizhou miniature Xiang pig , Guangxi Bama miniature pig and Landrace.There was no polymorphism obtained among or within three breeds of Chinese laboratory miniature pigs besides Landrace. It is concluded that the polymorphism of mtDNA D-loop within the three breeds of Chinese laboratory miniature pigs is poor , These methods cannot be used to distinguish among them , but it can be used to distinguish them from Landrace.     

三江白猪与其他5个品种猪的群体遗传关系的分析

应用100条RAPD引物对三江白猪及其杂交亲本长白猪和东北民猪,以及大约克夏、圣特西和法系大白猪进行亲缘关系的DNA多态分析。其中15条引物的扩增结果具有明显的多态,将其扩增结果进行聚类分析,发现三江白猪与长白猪（遗传距离d = 0.090）、与东北民猪（d = 0.175）的亲缘关系较近,与大约克夏(d = 0.263)、法系大白(d = 0.223)、圣特西猪(d = 0.580)的亲缘关系较远。Abstract: One hundred primers were used to analyze the polymorphism of kinship of Sanjiang White pig and its parents, Landrace and Dongbei Min pig, Yorkshire, French Hybrids(Fh 019 strain),French Yorkshire by RAPD. Amplification results of 15 primers showed obvious polymorphisms, which were analyzed by clustering. The results indicate that the genetic relationship of Sanjiang White Pig, and Landrace, and Dongbei Min pig are respectively closer(d=0.090,,0.175), and that of Sanjiang White pig and Yorkshive(d=0.263), French Yorkshire(d = 0.223), Fh 019strain (d = 0.580) are respectively further.     

赣中南花猪随机扩增多态DNA与群体遗传关系的研究

用RAPD技术检测了泰和冠朝猪、永丰藤田花猪、瑞金三花猪、兴国茶园猪、上犹花猪、万安花猪、乐安花猪等7个赣中南花猪地方类群基因组混合DNA的多态性,经80个随机引物扩增筛选,16个引物产生了共44个多态标记。遗传距离指数计算结果显示：泰和冠朝猪和瑞金三花猪亲缘关系最近,而兴国茶园猪与乐安花猪遗传距离最远。各类群的UPMGA和NJ聚类分析结果与采用生化、免疫和细胞遗传标记分析的结论不尽相同,在此基础上,结合现行分类方法,认为各受试猪群可暂时归并为赣中南花猪类型,但对泰和冠朝猪和万安花猪的品种归属性应作进一步分析。 Abstract: Random amplified polymorphic DNAs (RAPDs) were used to investigate bulked genomic DNA polymorphism of seven pig populations including Taihe Guanchao,Yongfeng Tengtian Spotted,Ruijin Spotted,Xingguo Chayuan,Shangyou Spotted,Wanan Spotted,and Lean Spotted pig.Eighty random primers were screened,of them,16 primers generated polymorphic markers with total number of 44.The genetic distance index matrix indicated that the genetic relationship between Taihe Gaunchao and Ruijin Spotted pig population was the closest,while that between Xingguo Chayuan and Laan Spotted pig population was the farthest.Our molecular phylogenetic trees constructed by UPMGA and NJ methods were in general consistent with that based on biochemical,immunogenetic and cellular genetic markers except for differences in some populations.It was inferred from this study that all pig populations examined could be temporary classified into one breed,however,the population genetic character of Taihe Guanchao and Wanan Spotted pig populations should be further evaluated.     

中外11个猪种A-FABP基因微卫星遗传变异的研究

利用微卫星技术对五指山猪、沂蒙黑猪、汉江黑猪、莱芜猪、北京黑猪、民猪、成华猪、内江猪、二花脸猪、巴马香猪和大白猪11个猪种共420头猪的脂肪细胞脂肪酸结合蛋白基因内含子的微卫星序列的遗传变异进行了研究。研究表明：（1）中国猪种在A-FABP基因微卫星基因座上有极高的的多态性,五指山猪的PIC最高（PIC=0.7904）并检测到11个等位基因,但北京黑猪仅检测到两个等位基因。（2）本实验中,只有北京黑猪、民猪、巴马香猪和大白猪A-FABP基因微卫星在群体中的分布达到平衡状态。（3）群体间的基因分化系数分析表明,中国猪种在该微卫星基因座上分化程度平均约为40.83%。Abstract：The genetic variations of microsatellites in intron 2 of the porcine adipocyte fatty acid-binding protein (A-FABP) genes were investigated in 420 pigs including Wuzhishan pig,Yimeng black pig,Hanjiang black pig,Laiwu pig,Beijing black pig,Min pig,Chenghua pig,Neijiang pig,Erhualian pig,Bama xiang pig and Large White pig.The results suggested as follows：（i）PIC of the Wuzhishan pig breed is the highest（0.7904） and 11 alleles were detected. Compared with Large White pig,Chinese pig breeds showed a great polymorphism of A-FABP microsatellites except Beijing black pig in which only 2 alleles were detected.（ii）Only Min pig, Bama xiang pig, Beijing black pig and Large White pig were in Hardy-Weinberg equilibrium. （iii）The analysis of genetic differentiation showed that the average value of the A-FABP gene differentiation of 10 Chinese pigs is about 40.83%.     

五种品系猪亲缘关系的RAPD分析

应用190个RAPD引物对贵州小型猪,巴马小型猪、西双版纳近交系小耳猪JB和JS亚系、荣昌猪Ⅰ系、长白猪的亲缘关系进行了初步分析,其中39个引物的扩增结果具有非常明显的品系间多态性,将其扩增结果以RAPDistance package Version 1?04程序进行分析,计算相对遗传距离指数1－F,再用NJ法进行聚类分析,构建系统树。结果表明,三种小型猪相互间亲缘关系较近,尤以贵州小型猪与巴马小型猪更近。荣昌猪Ⅰ系和长白猪关系较近,但与小型猪关系较远。 关键词：随机扩增多态DNA;小型猪;亲缘关系 The phylogenetic relationship of five species of pigs including Guizhou miniature pig,Bama pig,Xisuangbanna inbred pig,Rongchang Pig and Landrance was studied by RAPD analysis.Thirty-nine single polymorphic primers were selected out of 190 primers.The amplified fragments of thirty-nine primers were analysed by the RAPDistance package version 1.04 and the phylogenetic tree was constructed using NJ method.The results indicated as the following: three strains of miniature pigs have close phylogenetic relationship and the phylogenetic relationship between Guizhou miniature pig and Bama miniature pig was much closer.Landrance and Rongchang pig have close phylogenetic relationship too,but their phylogenetic relationship is far from the three strains of miniature pigs.     

三品系小型猪35个微卫星基因座的遗传学研究

利用35个微卫星基因座对中国三个品系的小型猪（贵州小型香猪、广西巴马小型猪、版纳小耳猪近交系）进行了遗传检测。计算出三个小型猪品系个体样本在35个微卫星基因座的纯合率,并对其进行t检验。计算出各品系的平均杂合度,多态信息含量（PIC）及品系间的遗传距离,并进行了系统聚类。结果表明三个品系的小型猪其基因纯合率均较高,其中版纳小耳猪近交系的基因纯合率最高;PIC和平均杂合度均较低;贵州小型香猪和广西巴马小型猪亲缘关系较近,并均与版纳小耳猪近交系的亲缘关系略远。 Abstract：The polymorphism of 35 microsatellites in the three miniature pig breeds in China(Guizhou miniature pig, Guangxi Bama miniature pig, Banna miniature pig inbred) was analysed. Rates of homozygote for 35 microsatellite loci in three miniature pig breeds were calculated,and t-test for them were performed. Mean heterozygosity and polymorphism information content(PIC) were calculated for all breeds, and genetic distances between these breeds were estimated. The dendrograms were obtained based on genetic distances. The results suggest that rates of homozygote in the three breeds are all high, and that is the highest in Banna miniature pig inbred. The results also suggest that polymorphism information content and mean heterozygosity in all the three breeds are low, and the genetic relationship between Guizhou miniature pig and Guangxi Bama miniature pig is closer than their relationship with Banna miniature pig inbred.     

混合线性模型下猪群间遗传联系的度量

采用关联指数(IC)和决定系数(CD)两种方法,度量混合线性模型遗传评估下 猪群体间的遗传关联性。结果表明,加拿大安大略省的大约克夏猪、长白猪、杜洛克猪和汉普夏猪4个主要品种群体间具有良好的遗传联系。CD法既组合了数据结构和信息量,又考虑了预测误差方差和遗传变异性,是一个选择判断遗传评估精度的好方法。 Abstract:Two criteria for the measures of genetic connectedness in mixed linear model of genetic evaluation are used:the degree of connectedness(IC)and the generalized coefficient of determination(CD).The results indicated that the data of four dominant swine breeds:Yorkshire,Landrace,Duroc and Hampshire in Ontario,Canada are well connected.The CD,which combines data structure and amount of information and also accounts for both prediction error variance and genetic variability,is a good method to select for judging the precision of a genetic evaluation.     

荧光素标记的庚型肝炎病毒基因探针的制备及应用

The sequences of HGV gene that had been reported were checked and analyzed. After comparison of HGV seqences from different virus strains, a gene fragment of 31 nucleotides (7121-7152 nt) served as a probe was labeled by fluorescein-N₆-dd ATP with terminal transferase. The specificity of HGV probe was tested by dot-blot hybridization with HCV RNA—related virus RNA, C.T DNA, HGV-RNA, and Southern-blot hybridization with RT-PCR product of HGV. Positive results obstained only from the HGV-RNA of positive sera that was confirmed by nested RT-PCR, all others were negative. The sensitivity of fluorescein-labeled probe was examined. The result showed that the fluorescein-labeled probe was able to check≥1.56 pg of target genome. The conformity of HGV-RNA detection using RT-PCR and HGV gene probe was 97.9%. The experiment suggested that fluorescein-labeled probe can be used in clinic detection and epidemic study.     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.