云南文山黄牛和迪庆黄牛遗传多样性的蛋白电泳研究

采用水平淀粉胶蛋白电泳技术分析了云南文山黄牛和迪庆黄牛的３３种血液蛋白及同工酶,共计３７个遗传座位,其中６个座位检测到多态性。文山黄牛的多态座位百分比Ｐ＝０．１３８９,平均杂合度Ｈ＝０．０６１０,迪庆黄牛Ｐ＝０．１６６７,Ｈ＝０．０６９１。通过Ｎｅｉ氏遗传距离计算,运用ＰＨＹＬＩＰ３．５Ｃ软件包中的“ＵＰＧＭＡ”,“ＣＯＮＴＭＬ”和“ＮＥＩＧＨＢＯＲ”法,结合前人报道的数据对１０个黄牛品种进行聚类分析。结果得出：文山黄牛可能主要起源于瘤牛（Ｂｏｓｉｎｄｉｃｕｓ）,迪庆黄牛可能主要起源于普通黄牛（Ｂｏｓｔａｕｒｕｓ）。我们的结果提示,云南黄牛可能是由当地人驯化的野生牛群与外来的驯化牛群相结合并适应当地气候和环境逐渐形成的,因而在蛋白水平上具有较为丰富的多样性。     

独龙牛遗传多样性及其种群遗传结构的等位酶分析

采用水平片淀粉凝胶电泳技术,进行３０头独牛牛４１种蛋白质共计４４个遗传座位的等位酶分析,只在Ｔｒ,Ｈｐ,Ａｍｙ,Ｅｓｔ等４个座位发现多态性。每个座位等基因的平均数、多态座位百分比和平均杂合度值分别为Ａ＝１．０９０９、Ｐ＝０．０６８２和Ｈ＝０．０２６２。贡山县和福贡县独龙牛群体从酶基因的角度上看遗传多样性贫乏,可能是分别由小种群引种而来,受到瓶颈效应的作用,并伴随着创立者事件的发生。我们结合独龙牛在     

中国猕猴类的蛋白多态性及其遗传分化关系

采用蛋白电泳分析技术研究了来源于我国和越南的猕猴属（Ｇｅｎ ｕｓＭａｃａｃａ）５个种（Ｍ．ｍｕｌａｔｔａ、Ｍ．ａｒｃｔｏｉｄｅｓ、Ｍ．ａｓｓａｍｅｎｓｉｓ、Ｍ．ｔｈｉｂｅｔａｎａ、Ｍ．ｆａｓｃｉｃｕｌａｒｉｓ）的蛋白多态性及其遗传分化关系。共分传座位３０个。在分析的１９只恒河猴中,３０个遗传座有位９个座位表现出多态性,多态座位百分比Ｐ＝０．３,平均等位基因数Ａ＝１．４,平均杂合度Ｈ＝０．０１０４５     

巴马小型猪群体遗传结构的随机扩增多态DNA分析

目的分析巴马小型猪的群体遗传结构。方法应用经过筛选的 ３１条引物对巴马小型猪个体基因组ＤＮＡ进行ＲＡＰＤ扩增,利用从ｉｎｔｅｒｎｅｔ网上下载的软件ＲＡＰＤ ｉｓｔａｎｃｅ Ｐａｃｋａｇｅ ｖｅｒ－ｓｉｏｎ１．０４软件对实验结果进行分析,计算不同个体间的相似系数。结果经ＲＡＰＤ反应,共扩增出２７５条带,其中多态性带８５条,多态性带频率在０～６６．７％之间,平均为２７．７％。不同个体猪拥有的ＲＡＰＤ条带具有差异,但拥有相同条带个体猪比率较高。该群体猪的相似系数（Ｆ）为０．９２８（０．７８～０．９７）,平均等位基因频率（ｑ）为０．７３２,最低平均杂合率（Ｈ）为０．２８６。结论巴马小型猪个体具有较好的遗传一致性和遗传稳定性。     

正常足月儿畸变产物耳声发射(DPOAEs)特性分析

研究提出正常足月儿畸变产物耳声发射（ＤＰＯＡＥｓ） 特性分析如下：１ ． ＤＰＯＡＥｓ 反应强度曲线,ＤＰ 图显示两个反应峰（ｆ２ ＝ １ ．６ 和５ ．０ ｋＨｚ） 和一个反应谷（ｆ２ ＝ ３ ．１ ～４ ．０ ｋＨｚ） ;２． ＤＰＯＡＥ 本底噪声及其特性,ｆ２ ＝ １．０ ｋＨｚ 其测试频率点（２ｆ１ －ｆ２） 本底噪声最高（Ｐ＜ ０ ．０５） ,ｆ２ ＝ ３ ．１ ,４．０ 和５ ．０ｋＨｚ ,测试频率点（２ｆ１ －ｆ２） 本底噪声较低（Ｐ＜ ０．０５） ;除ｆ２ ＝ ４．０ ｋＨｚ 外,ＤＰＯＡＥ 本底噪声与其反应强度均未呈现直线相关特性;３ ． ＤＰＯＡＥ ＳＮＲ 特性,ｆ２ ＝ １．０ ｋＨｚ 其ＳＮＲ 最小（Ｐ＝ ０．０００） ,ｆ２ ＝２ ．０ ｋＨｚ 其ＳＮＲ 最大（Ｐ０ ．００３） ;４． ＤＰＯＡＥ ＳＮＲ 和ＴＥＯＡＥ ＳＮＲ 相关特性,除１．０ ｋＨｚ 频段外,其余频段其二者间均有着非常显著的直线相关特性。     

糖基化对乙型肝炎表面抗原疫苗的影响

哺乳动物（ＣＨＯ）细胞表达的三种糖基化程度不同的乙肝表面抗原（ＨＢｓＡｇ）Ａ（含ＧＰ３０,ＧＰ２７,Ｐ２３）,Ｂ（含ＧＰ２７及Ｐ２３）,Ｃ（只有Ｐ２３）,在免疫原性、放置后抗原的稳定性以及对不同单克隆抗体亲和力等方面都有明显的差异;（１）免疫ＢＡＬＢ／Ｃ小鼠后血清中抗体工（ＥＤ５０）。平均结果为Ａ＝１：８０,Ｂ＝１：１１７,Ｃ＝１：１４,表明有糖基的ＨＢｓＡｇ疫苗对小鼠的免疫力明显高于无糖基的ＨＢ     

B链氨端去－肽（B1）羧端去五肽（B26—30）猪胰岛素的初步晶体学研究

应用悬滴气相扩散法在含６％氯化钠和１２．５％丙酮的柠檬酸缓冲体系中,获得可供Ｘ射线结构分析用的Ｂ链氨端去－肽（Ｂ１）羧端去五肽（Ｂ２６－３０）猪胰岛素单晶体。晶体属四方晶系,空间群为Ｐ４１２２或Ｐ４３２２,晶胞参数为：ａ＝ｂ＝３６．０Ａ,ｃ＝１２０．０Ａ,α＝β＝γ＝９０度。单位晶胞中每个结晶学不对称单位含有２个ＤｅｓＢ１－ＤＰＩ分子。     

B链氨端去－肽（B1）羧端去五肽（B26—30）猪胰岛素的初步晶体学研究

应用悬滴气相扩散法在含６％氯化纳和１２．５％丙酮的柠檬酸缓冲体系中,获得可供Ｘ射线结构分析用的Ｂ链氨端去一肽（Ｂ１）羧端去五肽（Ｂ２６—３０）猪胰岛素（ＤｅｓＢ１－ＤＰＩ）单晶体。晶体属四方晶系,空间群为Ｐ４１２２或Ｐ４３２２,晶胞参数为：ａ＝ｂ＝３６．０Ａ,ｃ＝１２０．０Ａ,ａ＝β＝γ＝９０°。单位晶胞中每个结晶学不对称单位含有２个ＤｅｓＢ１—ＤＰＩ分子。     

向日葵种质资源的随机扩增多态性ＤＮＡ（ＲＡＰＤ）研究

采用ＲＡＰＤ方法对我国２１个向日葵（Helianthus annuus L.)基因型和１１个国外引进向日葵因型进行分析,构建了它们的方图谱。从８０个随机引物中筛选出的２５个有效引物共产生１８８条ＤＮＡ片段,大小分布在０．２６－１．９８kb之间,其中１６４例带具有遗传多态性,约占总数的８７．２％,平均每个引物扩增的ＤＮＡ带数为７．５２条。３２个向日葵的遗传相似性分析表明,各基因型间的Ｎei氏相似性系数分布在０．３９８７．０８５３１之间,平均相似性系数为０．６２８１。１１个国外向日葵基因型的Ｎei氏相似性系数分布在０．７１３４－０．８５３１之间,平均相似性系数为０．７８２８,说明国外基因型之间的遗传基础比较狭窄。２１个国内向日葵基因型的Ｎei氏相似性系数分布在０．４７５０－０．８２０６,平均相似性系数为０．６４７８,说明国内向日葵基因型之间的遗传多态性较为丰富。通过非加权算术平均数聚类（ＵＰＧＭＡ）的方法,给制了３２个向日葵基因型之间的遗传关系树图。３２个向日葵基因明显地聚成Ａ、Ｂ两大类群。２１个国内向日葵基因型聚成了Ａ１、Ａ２两个亚类,Ａ１组包括：Ｘ１０、陕西向日葵、Ｄ－Ｓ１２、长岭向日葵６、黑２－Ｓ２－２、Ｔ－Ｃ０８、长岭葵花４、白葵３号、ＢＨ－１０、Ｊ－Ｓ－Ｂ１、张掖白子葵、Ｃ１０１－Ｓ４－３Ｓ４等１２个基因型;Ａ２组包括：ＬＫ３０５－Ｓ８、ＬＫ、ＣＳ－７、长岭葵花Ｓ２－Ｓ２、辉南７－Ｓ１－３、辉南、ＣＹ－ＸＸ１９－ＸＸ２、吉葵１１２、吉葵１１６等９个基因型。１１个国外向日葵基因型划分为Ｂ１、Ｂ２两个亚类,Ｂ１组包括ＬＧ１２０２８Ｑ、ＬＧ９０２３Ｒ、ＣＲＮ１４３、ＳＦ９９０３、ＳＦ９９０２、Ｓ－３３２２、ＳＨ３３２、ＳＨ４１、ＳＦＰ９００１、ＣＲＮ１４４５等１０个法国基因型;Ｂ２组织有来自美国的Ｇ１０１一个材料。     

高活力胰岛素研究：B3—Lys猪胰岛素的结晶及初步晶体学分析

用化学半合成方法制备的Ｂ３—Ｌｙｓ猪胰岛素（Ｂ３ＫＰＩ）具有与天然猪胰岛素相同的体内生物活力,但其体外生物活力及受体结合活力比天然猪胰岛素高。本文报道Ｂ３ＫＰＩ的晶体生长,初步晶体学分析及衍射数据与处理。Ｂ３ＫＰＩ晶体的衍射分辨率可以达到２．１Ａ（１Ａ＝０．１ｎｍ）,空间群为Ｒ３,晶胞参数为：ａＨ＝ｂＨ＝８２．２３ＡＣＨ＝３４．１７Ａ．收集了一套２．１Ａ分辨率的衍射数据,数据完全度为８１％,Ｒｍｅｒｇｅ为２．９％。     

Genetic Diversity of Chinese Native Pigs Inferred from Protein Electrophoresis

We examined protein polymorphism of 20 nativepig breeds in China and 3 introduced pig breeds. Thirtyloci have been investigated, among which six loci werefound to be polymorphic. Especially, the polymorphismof malate dehydrogenase (MDH), adenylate kinase(AK), and two new alleles of adenosine deaminase (ADA)had not been reported in domestic pigs and wild pigs.The percentage of polymorphic loci (P), the meanheterozygosity (H), and the mean number ofalleles (A) are 0.200, 0.065, and 1.300, respectively.The degree of genetic variability of Chinese pigs as awhole was higher than that of goats, lower than thatof cattle and horses, and similar to that ofsheep. Using the gene frequencies of the 30 loci, Nei'sgenetic distance among the 20 native breeds in China and3 introduced pig breeds was calculated by theformula of Nei. The program NEIGHBOR in PHYLIP3.5c was chosen to construct an UPGMA tree and a NJtree. Our results show that, of the total geneticvariation found in the native pig breeds in China, 31%(0.31) is ascribable to genetic differencesamong breeds. About 69% of the total genetic variationis found within breeds. Most breeds are in linkagedisequilibrium. The patterns of genetic similaritiesbetween the Chinese native pig breeds were notin agreement with the proposed pig type classification.     

Genetic Diversity of Yunnan Local Pig Breeds Inferred from Blood Protein Electrophoresis

Protein electrophoresis was used to examine theblood protein polymorphism in Yunnan local pig breeds,i.e., the Saba pig, Dahe pig, and Diannan small-ear pigbreeds. Of 38 genetic loci surveyed, 9 were found to be polymorphic. The percentage ofpolymorphic loci (P) varies from 0.1875 to 0.2121, andthe mean individual heterozygosity (H) varies from0.0712 to 0.1027 in three pig breeds. The resultsindicate that blood protein polymorphism in Yunnan pigbreeds is high. Yunnan local pig breeds have a wealth ofgenetic diversity at the level of blood proteins.     

Evaluation of genetic diversity in Chinese indigenous chicken breeds using microsatellite markers

China is rich in chicken genetic resources, and many indigenous breeds can be found throughout the country. Due to poor productive ability, some of them are threatened by the commercial varieties from domestic and foreign breeding companies. In a large-scale investigation into the current status of Chinese poultry genetic resources, 78 indigenous chicken breeds were surveyed and their blood samples collected. The genomes of these chickens were screened using microsatellite analysis. A total of 2740 individuals were genotyped for 27 microsatellite markers on 13 chromosomes. The number of alleles of the 27 markers ranged from 6 to 51 per locus with a mean of 18.74. Heterozygosity (H) values of the 78 chicken breeds were all more than 0.5. The average H value (0.622) and polymorphism information content (PIC, 0.573) of these breeds suggested that the Chinese indigenous chickens possessed more genetic diversity than that reported in many other countries. The fixation coefficients of subpopulations within the total population (F _ST) for the 27 loci varied from 0.065 (LEI0166) to 0.209 (MCW0078), with a mean of 0.106. For all detected microsatellite loci, only one (LEI0194) deviated from Hardy-Weinberg equilibrium (HWE) across all the populations. As genetic drift or non-random mating can occur in small populations, breeds kept on conservation farms such as Langshan chicken generally had lower H values, while those kept on large populations within conservation regions possessed higher polymorphisms. The high genetic diversity in Chinese indigenous breeds is in agreement with great phenotypic variation of these breeds. Using Nei’s genetic distance and the Neighbor-Joining method, the indigenous Chinese chickens were classified into six categories that were generally consistent with their geographic distributions. The molecular information of genetic diversity will play an important role in conservation, supervision, and utilization of the chicken resources.     

Assessment of genetic diversity of Korean native pig (Sus scrofa) using AFLP markers

In order to assess the genetic diversity and genetic relationships among the six commercial pig breeds including the Korean native pig, we performed an amplified fragment length polymorphism (AFLP) analysis. Applying the three EcoRI/TagI primer combinations to 54 individual pig samples out of six breeds, a total of 186 AFLP bands were generated, 67 (36%) of which were identified as polymorphic bands. From these polymorphic bands, the three estimates (percentage of polymorphic loci, Neis heterozygosity and Shannon index) of genetic diversity, G(ST) estimates, Neis unbiased genetic distance and two indices of genetic similarity were calculated. From all the calculations of genetic diversity, the lowest genetic diversity was exhibited in the Korean native pig, and the highest in the Chinese Yanbian pig. Given the mean G(ST) value (G(ST) = 0.390) across all pigs examined, levels of apparent breed subdivision were considerable. A UPGMA tree of individuals based on Jaccards similarity index showed that the Korean native pig formed a distinct cluster from the other five pigs. In addition, the tree displayed that all the individuals except for six individuals were grouped into their breeds. Principal component analysis based on the binary data matrix of either presence or absence confirmed the distinctness of the Korean native pig from the other pigs. Our results indicate that the Korean native pig has a low level of genetic diversity and is distinct from the five pig breeds, confirming the results from previous microsatellite data. The findings also suggest that AFLP analysis may be a valuable tool for revealing genetic relationships and genetic diversity among different pig breeds.     

Evaluation of genetic diversity in Chinese indigenous chicken breeds using microsatellite markers

China is regarded as one of the domestication cen-ters for chickens and archaeological studies provided evidence of chicken domestication in northern Chinaas early as 6000 BC[1]. At present, China has the larg-est chicken population in the world, represen…     

Genetic diversity and population structure of six Chinese indigenous pig breeds in the Taihu Lake region revealed by sequencing data

The Chinese indigenous pig breeds in the Taihu Lake region are the most prolific pig breeds in the world. In this study, we investigated the genetic diversity and population structure of six breeds, including Meishan, Erhualian, Mi, Fengjing, Shawutou and Jiaxing Black, in this region using whole‐genome SNP data. A high SNP with proportions of polymorphic markers ranging from 0.925 to 0.995 was exhibited by the Chinese indigenous pigs in the Taihu Lake region. The allelic richness and expected heterozygosity also were calculated and indicated that the genetic diversity of the Meishan breed was the greatest, whereas that of the Fengjing breed was the lowest. The genetic differentiation, as indicated by the fixation index, exhibited an overall mean of 0.149. Both neighbor‐joining tree and principal components analysis were able to distinguish the breeds from each other, but structure analysis indicated that the Mi and Erhualian breeds exhibited similar major signals of admixture. With this genome‐wide comprehensive survey of the genetic diversity and population structure of the indigenous Chinese pigs in the Taihu Lake region, we confirmed the rationality of the current breed classification of the pigs in this region.     

The phylogeny of Chinese indigenous pig breeds inferred from microsatellite markers

A genetic study of 32 local Chinese, three foreign pig breeds [Duroc (DU), Landrace and Yorkshire], and two types of wild boar (Hainan and Dongbei wild boar) based on 34 microsatellite loci was carried out to clarify the phylogeny of Chinese indigenous pig breeds. The allele frequencies, effective numbers of alleles, and the average heterozygosity within populations were calculated. The results showed that the genetic variability of the Lingao pig was the largest, while the Jiaxing pig was the lowest. The greatest distance between domestic pigs was found between Shanggao and DU pig and the shortest was found between Wuzhishan and Lingao pig, respectively. A neighbour-joining tree constructed from Modified Cavalli-Sforza genetic distances divided Chinese pigs into two clusters; four subclusters were also identified. Our results only partly agree with the traditional types of classification and also provide a new relationship among Chinese local pig breeds. Our data also confirmed that Chinese pig breeds have a different origin from European/American breeds and can be utilized in programmes that aim to maintain Chinese indigenous pig breeds.     

Evaluation of the genetic diversity of Laiwu pigs using twenty-seven microsatellite markers

The Laiwu pig, an indigenous pig breed known for extremely high intramuscular fat content, is a well-preserved ancient breed due to long-term natural and artificial selections. In this study, using 27 microsatellite markers jointly recommended by the International Society of Animal Genetics (ISAG) and the Food and Agriculture Organization (FAO), we investigated the genetic diversity of the Laiwu pig breed. The genetic diversity of Laiwu pigs is dramatically low, with the observed heterozygosity ranging from 0.067 to 0.767. Among the 27 microsatellite markers, 10 were high polymorphic loci, 10 were moderate polymorphic loci, six were low polymorphic loci, and no polymorphism was detected at one locus (IGFI). Further analyses with the 10 high polymorphic loci and five moderate polymorphic loci revealed that the Laiwu pig breed was inbred and heterozygous deficient to some extent, but not severely, and that the Laiwu pigs were relatively pure, with almost no hybridization with other breeds. Two subgroups of the current 13 Laiwu pig pedigrees were identified. These results suggest that the Laiwu pig breed has a low diversity and a conservation program must be developed to preserve the “Laiwu pig” gene pool.     

Genetic diversity and differentiation of Chinese domestic buffalo based on 30 microsatellite markers

To determine genetic diversity and evolutionary relationships among Chinese buffalo populations, 18 indigenous swamp buffalo populations and two introduced river buffalo breeds were genotyped for 30 microsatellite loci. The mean number of alleles across the 30 loci was 8.13, and the expected heterozygosity ranged from 0.517 (Yanjin) to 0.609 (Dehong). Although there was abundant genetic variation, genetic differentiation between Chinese buffalo populations was low, with only 2.8% of the total genetic variance among populations. The genetic differentiation pattern and genetic relationships among Chinese buffalo populations were consistent with their geographic distribution. The Dehong population was discerned as a distinct indigenous population, but suffered genetic admixture with river buffalo. The remaining populations were divided into four major clusters, i.e. the Upper and Middle Reaches of Yangtze Valley cluster (Guizhou, Guizhoubai, Yanjin, Fuling, Enshi and Jianghan), the Lower Reaches of Yangtze Valley cluster (Haizi, Shanqu and Dongliu), the South China cluster (Fuan and Xinfeng) and the Southwest China cluster (Xinglong, Xilin, Diandongnan and Dechang).     

利用微卫星标记分析我国13个地方灰羽鹅品种的遗传多样性

我国地方鹅品种具有很多优良性状,但长期以来由于没有形成科学的选育制度和培育方法,品种的遗传多样性没有得到完整保存。为了深入了解我国地方鹅品种的遗传结构,使之得到更好的保护和利用,作者选用31个多态性较高的微卫星标记(其中19个是首次用磁珠富集法从AFLP片段中分离),检测了我国13个地方灰羽鹅(An-sercygnnoides)品种的遗传多样性。利用等位基因频率计算出各群体的平均遗传杂合度(H)、多态信息含量(PIC)和群体间的遗传距离(DA)。结果表明:13个地方灰羽鹅品种中,平均多态信息含量为0.323–0.398,平均杂合度为0.4985–0.6727,各品种的杂合度都较高,最高的是狮头鹅(0.6727),最低的是雁鹅(0.4985)。用UPGMA法进行聚类分析的结果显示,13个品种被聚为4类:丰城灰鹅、武冈铜鹅、兴国灰鹅、狮头鹅、乌棕鹅、阳江鹅、马冈鹅、钢鹅、雁鹅聚为第1类;伊犁鹅自聚为第2类;长乐鹅、右江鹅聚为第3类;永康灰鹅自聚为第4类。本研究为鹅遗传育种提供了参考资料。