中华拟德氏吸虫与日本血吸虫的RAPD遗传标志分析

中华拟德氏吸虫(Paradeontacylix sinensis)是冷血吸虫,日本血吸虫(Schistosoma japonicum)是温血吸虫的重要代表。分别对中华拟德氏吸虫和日本血吸虫成虫的全基因组进行随机扩增多态性研究,筛选出31种随机引物,对2种成虫扩增得到189条多态性基因片断,S6、S46对中华拟德氏吸虫全基因组有特异性,分别扩增出一条大小约399bp和695bp的特异性强条带;引物S2对日本血吸虫基因组有特异性,只扩增1条1670bp的特异性强条带。多数引物对中华拟德氏吸虫和日本血吸虫成虫全基因组扩增出大小相似的条带,推测中华拟德氏吸虫与日本血吸虫的亲缘关系密切。     

Genetic diversity among Escherichia coli O149:K91 strains isolated from pigs with diarrhoea determined by randomly amplified polymorphic DNA analysis.

The genetic relatedness among 72 Escherichia coli strains of serotype O149:K91 isolated from pigs with diarrhoea was investigated by randomly amplified polymorphic DNA (RAPD) analysis. Fimbrial and toxic virulence markers of the isolates were also tested. Amplification with primer 1254 resulted in three different RAPD types whereas primer 1290 generated one RAPD profile only. Based on the RAPD and fimbrial/toxin types the strains were classified into five distinct groups.     

Evaluation of canine Bordetella bronchiseptica isolates using randomly amplified polymorphic DNA fingerprinting and ribotyping

Bordetella bronchiseptica is a respiratory tract pathogen in a variety of species. Previous studies suggest little genetic variation among canine B. bronchiseptica isolates. The degree of genetic diversity in 26 canine B. bronchiseptica strains was evaluated using randomly amplified polymorphic DNA (RAPD) fingerprinting and ribotyping. Strains evaluated include historic reference strains (N=3). vaccine strains (N=5) and clinical isolates (N=18). RAPD fingerprinting with the 10-nucleotide primer OPA-4 resulted in four distinct fingerprint patterns. RAPD fingerprinting consistently separated four previously characterized electromorphotype (EMT) 6 strains into two fingerprint types. Ribotyping, using the restriction endonuclease PvuI, resulted in six distinct ribotypes. With the exception of vaccine strains, considerable genetic diversity exists in the canine B. bronchiseptica isolates examined. These findings indicate the genetic variability within canine strains of B. bronchiseptica is greater than appreciated previously. Additionally, OPA-4 RAPD fingerprinting and PvuI ribotyping will be useful tools in epidemiologic studies of canine B. bronchiseptica isolates.     

皖北猪的随机扩增多态DNA研究

为保护和开发地方品种猪的遗传资源,采用随机扩增多态DNA(RAPD)法分析皖北猪个体之间的遗传多样性。试验方案采用L(35)正交法,优化RAPD反应条件;筛选7条特异引物对皖北猪进行RAPD分析,计算带谱相似率、平均杂合度和遗传相似度,UPGMA法进行聚类分析。试验结果表明,优化的皖北猪RAPD反应体系为Mg2+浓度3 mmol/L,Taq酶1.50 U,dNTPs浓度0.40 mmol/L,引物浓度0.50μmol/L,模板浓度1 mg/L。皖北猪群处于亲缘关系不清的遗传非均衡状态。     

应用随机扩增多态性进行山羊致病性大肠杆菌的分子多态性分析

为了解山羊致病性大肠杆菌广西分离株的分子多态性,应用随机扩增多态性方法(RAPD)对山羊致病性大肠杆菌进行分型研究.从8条随机引物中筛选出4条能在10株大肠杆菌中具有较好多态性扩增的随机引物,4条随机引物共扩增出18条DNA片段,10个菌株无共有带谱,显示出良好的扩增多态性.菌株Nx31与Nx32曾被认为是同一菌株的两次分离,但是在RAPD分析中,两株细菌的带谱存在明显的差别,表明RAPD比传统的血清学分型具有更高的分辨性.     

Complementary randomly amplified polymorphic DNA (RAPD) analysis patterns and primer sets to differentiate Mycoplasma gallisepticum strains.

Randomly amplified polymorphic DNA (RAPD) analysis was used to differentiate 7 strains of Mycoplasma gallisepticum. Six commercially available primers or primer combinations were screened for their ability to differentiate vaccine and type strains. Although major and minor bands were produced with each primer, many of the primers were unsuitable for strain differentiation. The use of primer 6 and combined primers 3 and 4 resulted in complementary RAPD banding patterns for each M. gallisepticum strain. Eleven different isolates representing 7 different strains were segregated into 7 different patterns, corresponding to the 7 strains.     

用RAPD标记分析高羊茅的遗传多样性

采用RAPD分子标记技术对从国外引进的15个高羊茅品种的遗传多样性进行了研究。从60个随机引物中筛选出12个有效引物,它们共扩增出85条DNA带,其中59条为多态性带,占69.41%,平均每个引物扩增出多态性带4.92条;利用NTSYS-PC软件计算出的不同品种间Jaccard遗传相似系数(GS)变化范围较大,为0.373~0.932;根据得到的遗传相似性矩阵进行非加权组法(UPGMA)聚类分析,建立高羊茅品种的分子系统树状图;以相似系数0.68为标准,可将所有品种分为3类,品种翠丽和贝克各自聚为一类,其余13个品种聚成一类。     

Genomic fingerprinting of pigeon Streptococcus gallolyticus strains of different virulence by randomly amplified polymorphic DNA (RAPD) analysis

Randomly amplified polymorphic DNA (RAPD) analysis was performed on 95 pigeon S. gallolyticus strains of different virulence and belonging to different biotypes and different culture supernatant phenotypes as determined by SDS-PAGE. Four distinct RAPD patterns, designated A, B, C and D, were distinguished using primer OPM6 (5'CTGGGCAACT). All 76 strains generating RAPD pattern A or B were designated highly virulent on the basis of their SDS-PAGE pattern. Five of seven strains generating RAPD pattern C and 11 of 12 strains generating RAPD pattern D belonged to the moderately virulent and low virulent culture supernatant phenotype groups, respectively. Only one RAPD group C strain belonged to a highly virulent culture supernatant phenotype group. There was a correlation between biotype and RAPD patterns. These findings indicate that there is a high correlation between RAPD pattern and virulence for pigeons. Therefore, RAPD typing seems a rapid, reliable method to distinguish pigeon S. gallolyticus strains of high, moderate and low virulence.     

Differentiation between high and low virulence Staphylococcus aureus strains from rabbits by randomly amplified polymorphic DNA (RAPD) analysis

Randomly Amplified Polymorphic DNA (RAPD) typing was performed on 53 rabbit Staphylococcus aureus strains. Twenty-three strains isolated in 13 different rabbitries with chronic problems of staphylococcosis, showed the same RAPD banding pattern. Twenty of these strains belonged to the 'mixed CV-C' biotype and to the phage-type 3A/3C/55/71, previously described to be highly virulent in rabbits, and three strains belonged to other biotypes or phage-types. None of the strains isolated from rabbitries without chronic problems of staphylococcosis showed this specific RAPD pattern. RAPD analysis can be used as a rapid and reliable test method to differentiate between the characteristic genotype corresponding to high virulence and other S. aureus strains from rabbits. This is useful for the diagnosis and prevention of the introduction of these highly virulent strains in industrial rabbitries.     

Phenotypic characterization and random amplified polymorphic DNA (RAPD) analysis of Pasteurella multocida isolated from Korean pigs

Pasteurella multocida causes various respiratory disease symptoms in pigs, including atrophic rhinitis and pneumonia. In the present study, 69 strains of P. multocida were isolated from 443 pigs with respiratory clinical symptoms at 182 farms located throughout South Korea from 2009 to 2010. A multiplex capsular PCR typing assay revealed that 69 strains of P. multocida isolated in this study had the biosynthetic locus of the capsules of either serogroup A (47 strains, 68.1%) or serogroup D (22 strains, 31.9%). The 22 strains positive for serogroup D-specific primers were divided into four clusters and the 47 strains positive for serogroup A-specific primers were divided into 12 clusters according to the results of Random Amplified Polymorphic DNA (RAPD) analysis. P. multocida strains in the present study were susceptible to most of the antimicrobial agents used. An analysis of antimicrobial resistance and virulence gene pattern combined with RAPD indicated that a certain P. multocida strain appeared to be genetically identical, implying the persistence of the strain within a single farm.     

RAPD技术及其在中药DNA鉴定中的应用

DNA分子的多态性检测是进行基因组研究的基础。继 RFL P(Restriction Fragment LengthPolymorphism)和 DNA指纹 (DNA fingerprinting)技术后 ,1990年 Williams等人〔1〕第一次用随机引物扩增的方法寻找多态性 DNA片段 ,并用做分子标记 ,将此方法命名为 RAPD (Random AmplifiedP     

Genetic parameters for production traits and measures of residual feed intake in Duroc and Landrace pigs

Data on 380 Duroc boars from seven generations, and 1026 Landrace pigs (341 boars and 685 gilts) from six generations were used to estimate genetic parameters for daily gain (DG), backfat thickness (BF), metabolic weight (MWT), daily feed intake (FI), feed conversion ratio (FCR) and residual feed intake (RFI). Two measures of RFI were estimated as the difference between actual feed intake and that predicted from models that included initial test age and weight and DG (RFI1); and initial test age and weight, DG and BF (RFI2). Heritability estimates for DG, MWT and FI were moderate for both breeds. BF estimates were high for both the breeds. The measures of feed efficiency (FCR and RFI) were moderately heritable. Genetic correlations of BF with measures of RFI were stronger when BF was not included in the estimation of RFI (0.40 and 0.46 for Duroc and Landrace, respectively (for RFI1), compared with 0.05 and 0.06 for Duroc and Landrace, respectively (for RFI2)). Genetic correlations of MWT with measures of RFI were all negative and low. Genetic and phenotypic correlations between DG and measures of RFI were close to zero, which indicated that selection for reduced RFI could be made without adversely affecting DG. BF should also decrease, and MWT should increase under selection for reduced RFI. The reduction in BF would depend on the measure of RFI used.     

藏系绵羊随机扩增多态性DNA最佳反应体系的研究

以高原型藏羊的基因组DNA为模板，通过对RAPD反应体系中的各种参数进行优化试验，建立了适宜于藏系绵羊RAPD分析的最佳反应体系：在PE2400型DNA扩增仪的25μL反应体系中，模板DNA的量为60－120ng，引物量为4－8pmol，Mg^2 浓度为2.0mM，Taq DNA聚合酶为1－2.5U,dNTP浓度为150－200μM;94℃预变性3分钟后35次循环的参数设定为：94℃1分钟，36℃1分钟，72℃2.5分钟，最后72℃延伸10分钟，利用这些条件对藏系绵羊的基因组DNA进行RAPD分析，其结果的重复性和可靠性大为提高。     

对剩余采食量和生长性状进行7个世代选择的遗传参数估计

剩余饲料量(Residual Feed Intake,RFI)是动物实际的饲料摄入量减去预期动物用于生长的饲料消耗量后得到的一个差值。利用1 642头(380头公猪、868头后备母猪和394头阉猪)并历经7代杜洛克猪测试所得到的数据,对剩余采食量(RFI)、采食量利用(RFI)、采食量(Feed Intake,FI)、日增重(Average Daily Gain,ADG)、背膘(Back Fat,BF)和眼肌面积(Loin Eye Area,LEA)进行遗传参数的估(Feed Intake,FI)、日增重进行遗传参数的估计和相关计算。采用4种模型评估RFI。RFI1:包括了入试期、体重和ADG;RFI2:包括了入试期、体重、ADG和BF;RFI3:计和相关计算。采用BF;RFI3:包括了入试期、体重、ADG和LEA;RFI4:包括了入试期、体重、ADG、BF和LEA。然后再用最大似然法(REML法)对上述包括了入试期、体重、ADG对上述的动物模型进行遗传参数的估计。RFI的遗传力处于中等(范围在0.22~0.38)。相应对于FI,ADG和LEA的遗传力也处于中等(范围在0.45~0.49),但BF的遗传力较高(0.72)。FI与ADG的遗传相关较高,为0.84;FI与BF的遗传相关也较高,为0.67。LEA围在0.67。LEA与FI的遗传相关-0.42,LEA与ADG的遗传相关为-0.11,LEA与BF的遗传相关为-0.44。当BF没有包含在RFI的模型中时,BF与RFI的遗传相关较高(BF与RFI1的遗传相关为0.77,与RFI3的遗传相关为0.76);而相比之下,BF与RFI2的遗传相关为0.11;与RFI4的遗传相关为0.07。LEA与RFI的遗传相关都为负(范围为-0.30到-0.60)。与-0.60)。对ADG、LEA和肌内脂肪含量进行选择,当对RFI进行评估时,会得到比较小但与意愿相符合的遗传进展。4个RFI之间对之间的表型相关为0,遗传相关较低(0.17~0.23)。FI与所有的RFI都存在强相关,不管是遗传相关(0.56~0.77)还是表型相关的表型相关为还是表型相关(0.56~0.66)。结果显示:对RFI进行选择也许会导致FI的降低,BF也会变薄,LEA会增加。至于BF或LEA的改变量将相对(0.56的改变量将相对取决于RFI是否对BF进行?     

Genetic parameters for feed efficiency traits and their relationships with growth and carcass traits in Duroc pigs

Genetic parameters for feed efficiency traits of 380 boars and growth and carcass traits of 1642 pigs (380 boars, 868 gilts and 394 barrows) in seven generations of Duroc population were estimated. Feed efficiency traits included the feed conversion ratio (FCR), and nutritional (RFI(nut)), phenotypic (RFI(phe)) and genetic (RFI(gen)) residual feed intake. Growth and carcass traits were the age to reach 105-kg body weight (A105), loin eye muscle area (EMA), backfat (BF), intra-muscular fat (IMF) and meat tenderness. The mean values for RFI(phe) and RFI(gen) were close to zero and for RFI(nut) was negative. All the measures of feed efficiency were moderately heritable (h(2) = 0.31, 0.38, 0.40 and 0.27 for RFI(nut), RFI(phe), RFI(gen) and FCR respectively). The heritabilities for all growth and carcass traits were moderate (ranged from 0.37 to 0.45), except for BF, which was high (0.72). The genetic correlations of RFI(phe) and RFI(gen) with A105 were positive and high. Measures of RFI were correlated negatively with EMA. BF was more strongly correlated with measures of RFI (r(g) > or = 0.73) than with FCR (r(g) = 0.52). Selection for daily gain, EMA, BF and IMF caused favourable genetic changes in feed efficiency traits. Results of this study indicate that selection against either RFI(phe) or RFI(gen) would give a similar correlated response in carcass traits.     

Pulsed-field gel electrophoresis genotyping of Salmonella gallinarum and comparison with random amplified polymorphic DNA

Salmonella gallinarum is gram-negative bacteria that cause fowl typhoid (FT) in chickens. Since the first outbreak of FT reported in 1992 in Korea, it has widely spread throughout the country. Today, FT is one of the most devastating diseases of poultry. The aim of the present study was to ascertain a genetic relationship among S. gallinarum isolates collected from different regions of Korea over a 10-year period. We examined a total of 38 isolates of S. gallinarum obtained in 29 regions of Korea from 1992 to 2001 including the 9R vaccine strain and the standard strain of S. gallinarum (ATCC 9184). The PFGE profiles produced 12 different patterns with the XbaI-digestion and 11 different patterns with the SpeI-digestion. The RAPD using URP-6 primers showed eight different genotypes with the same Salmonella isolates. The PFGE patterns of the 9R vaccine strain and ATCC 9184 of S. gallinarum were different from the identical type A, the most common genotype among field isolates in our study. In conclusion, a low genetic heterogeneity was observed among Korean S. gallinarum isolates. In addition, PFGE appeared to be a more accurate and reproducible method for genotyping of S. gallinarum isolates than RAPD.     

Random amplified polymorphic DNA (RAPD) analysis of Mycobacterium tuberculosis strains in India

The usefulness of random amplification of polymorphic DNA (RAPD) analysis for typing Indian strains of M. tuberculosis was investigated. M. tuberculosis H37Rv, M. tuberculosis DT and 42 clinical isolates of M. tuberculosis were subjected to RAPD-PCR using 7 random decamer primers. All 7 primers were found to be differentiated and produced specific RAPD profiles. The polymorphic amplicons served as RAPD markers for M. tuberculosis. The dendrograms, obtained by different primers, showed the discriminatory ability of the primers. RAPD analysis provided a rapid and easy means of identifying polymorphism in M. tuberculosis isolates, and it was found to be a valuable alternative epidemiological tool. In addition, the results of the present study showed heterogeneity in the M. tuberculosis strains in the population studied.     

Genetic correlation between serum insulin-like growth factor-1 concentration and performance and meat quality traits in Duroc pigs

This study was intended to examine whether serum IGF-I concentration is appropriate for use as a physiological predictor for genetic improvement of meat production and meat quality traits in pigs. Heritabilities and genetic correlations were estimated for these traits. The Duroc breed used in this study was selected for seven generations for average daily BW gain (DG) from 30 to 105 kg of BW, loin-eye muscle area (EM), backfat thickness (BF), and intramuscular fat (IMF) content. Serum IGF-I concentration of boars and gilts at the fourth generation of selection and that of boars, gilts, and barrows from the fifth to seventh generations of selection were measured at 8 wk (IGFI-8W) for 832 animals and again at the time they reached 105 kg of BW (IGFI-105KG) for 834 animals. A multivariate REML procedure was used to estimate genetic parameters with a model incorporating generation of selection, sex, common environmental effect of litter, and individual additive genetic effects. Heritability estimates for IGFI-8W and IGFI-105KG were 0.23 +/- 0.02 and 0.26 +/- 0.03, respectively. The estimates of common environmental effect for IGFI-8W and IGFI-105KG were 0.20 +/- 0.02 and 0.03 +/- 0.01, respectively. Positive genetic correlations were estimated between IGFI-8W and DG (0.26 +/- 0.08), EM (0.22 +/- 0.10), and IMF (0.32 +/- 0.10). Moreover, the positive genetic correlation between IGFI-105KG and EM was 0.42 +/- 0.08. These results indicate that serum IGF-I concentration at an early stage of growth was effective for prediction of IMF, but it was not a reliable physiological predictor of genetic merit of meat production traits.