高丹草遗传图谱构建及农艺性状基因定位研究

本文以高丹草(So rghumSudan grass)(314A×ZKSD)的F_2:3家系作为构图群体, 构建了高丹草AFLP和RAPD遗传图谱。将248个F_2:3家系按随机区组重复3次的田间设计在两个试验点种植, 考察了包括产量在内的10个重要农艺性状。利用复合区间作图法分析10个性状的数量性状基因座位(QTL)以及基因效应值和基因作用方式, 并采用遗传分离分析法进行多世代联合分析, 主要结果如下。     

Scrapie resistance alleles are not associated with lower prolificity in Rasa Aragonesa sheep

Scrapie is a prion disease characterised by the accumulation of the pathological associated form of cellular prion protein (PrP(SC)) in the central nervous system. Susceptibility to scrapie is associated with polymorphism in the ovine prion protein (PrP) gene. The European Union has implemented scrapie control programs, relying on selective breeding for scrapie resistance; the use of ARR-carrier and the exclusion of VRQ-carrier were recommended. In this study, 4323 individuals from Rasa Aragonesa Sheep breed were genotyped for the PrP gene and the individual estimated breeding values (EBV) for prolificity were calculated. Most represented PrP alleles do not work against prolificity. Only a significant association between VRQ/VRQ genotype and a lower EBV was observed (p = 0.027, eta2 = 0.002). Therefore, avoiding reproduction of VRQ/VRQ individuals would not cause negative effect regarding prolificity.     

半胱亚磺酸脱羧酶(CSD)基因在大鼠睾丸组织中的表达及其序列分析

试验根据大鼠肝脏牛磺酸生物合成关键酶——半胱亚磺酸脱羧酶(CSD)基因已知核苷酸序列,设计了1对特异的寡核苷酸引物,对提取的睾丸组织总RNA进行RT-PCR扩增,扩增出468 bp的DNA片段,在国际上首次证明CSD基因在大鼠睾丸组织中存在表达,即牛磺酸可在大鼠睾丸组织中生物合成。测定了扩增的睾丸组织CSD基因核苷酸序列,并与已知肝脏CSD基因核苷酸序列及其对应的氨基酸序列作了同源性分析。结果表明:睾丸组织CSD基因核苷酸序列同已知肝脏CSD基因核苷酸序列的同源性为99.8%,其对应氨基酸序列的同源性为100%。     

扁蓿豆遗传多样性研究初探

论述了扁蓿豆遗传多样性研究的依据,利用不同生境条件下生长的野生和栽培品种,采用形态标记、细胞标记、生化标记和分子标记的方法多方面地对扁蓿豆进行遗传多样性的研究。     

中俄猪16个遗传标记基因位点的多态与繁殖性状的相关研究

检测了俄罗斯大白猪及萍乡华中二头乌猪１６个遗传标记基因位点的多态性,分析了二个猪种群体间在上述１６个基因位点上的遗传变异,探讨了所测遗传标记基因位点与种猪繁殖性状的相关性。同时,对两个猪种母猪体内抗红细胞抗原“天然抗体”的存在种类及效价与其繁殖性状的相关性进行了研究;     

Practical aspects of genetic management of small populations – The Olkuska sheep example

Abstract

The study aimed to identify important factors threatening genetic diversity within small sheep populations, and to propose sustainable management strategies. Demographic evolution, genetic diversity and current genetic management practices were analyzed in a population of Polish Olkuska sheep. Unbalanced contributions of individual rams to the next generation were observed due to differences in the period of their utilization, the number of progeny and the number of daughters used in reproduction. Matings between closely related animals were common. Pedigree completeness was satisfactory and inbreeding coefficient was high, 11.71% for lambs born in 2012. Relatively high values of potential and realized coancestries within flocks showed the importance of keeping more than one ram. The genetic conservation index and average relatedness coefficient, and their comparison between regions was helpful in considering management options. The information on genetic relationships and genetic conservation parameters of rams should be provided to breeders with simple interpretations.     

青海血蜱cDNA表达文库的免疫学筛选和阳性克隆的鉴定

为获得抗青海血蜱免疫原基因，用免抗青海血蜱差异蛋白阳性血清和兔抗青海血蜱唾液蛋白阳性血清对青海血蜱cDNA表达文库进行了免疫学筛选，经过初筛和复筛共获得58个阳性信号。用所得阳性噬菌体转染宿主菌BM25．8使之自动亚克隆为重组质粒，用此亚克隆质粒转化宿主菌JM109并从中提取重组质粒进行PCR、酶切和测序分析。序列分析表明：共获得新cDNA序列21个。将前5个cDNA登录GenBank／ncbi，获取登录号。所获阳性克隆为青海血蜱保护性抗原的筛选奠定了基础。     

美洲型与欧洲型PRRSV特异N蛋白抗原表位基因表达及其产物的纯化

对原核中以包涵体形式高效表达的美洲型与欧洲型PRRSV特异N蛋白抗原表位区段进行了变性、复性与纯化研究。将美洲型PRRSV重组质粒pGEX-6P-1-N转化入E.coliBL21(DE3)细胞中,成功表达了重组蛋白GST-N,超声裂解复性后目的蛋白经GSTrap FF纯化试剂盒纯化后,纯度可达90%以上。将欧洲型PRRSV重组质粒PQE30-N转化入E.coliM15细胞中表达了重组蛋白His-N。超声裂解后目的蛋白经HisTrapHP蛋白纯化试剂盒纯化后,纯度可达95%以上。Western blot结果表明,两种蛋白分别被美洲型和欧洲型PRRSV阳性血清识别。纯化出的大量美洲型与欧洲型PRRSV特异N蛋白抗原表位区段,为PRRSV的鉴别诊断提供了抗原基础。     

H5N1亚型禽流感病毒NS1基因的原核表达及其ELISA检测方法的建立

采用RT-PCR技术扩增了禽流感病毒（AIV）A／Goose／HLJ／p46／2003（H5N1）的NSl基因，将其克隆于融合蛋白表达载体pMAL-c2X上，转化DH5α大肠埃希氏菌感受态细胞，经BamHⅠ和HindⅢ双酶切鉴定及序列分析，表明筛选到了重组质粒pc2X-NS1。SDS-PAGE电泳结果显示，重组质粒转化TB1大肠埃希氏菌后，经0．3mmol／L的IPTG诱导，融合蛋白MBP-NS1得到大量表达，融合蛋白以可溶形式存在，分子质量约为67ku。Western-blotting检测结果表明，融合蛋白MBP-NS1能够与H5N1亚型AIV活病毒感染康复鸭血清发生特异性反应，而不能够与H5N1亚型AIV灭活疫苗免疫鸭血清发生反应。试验初步建立了以纯化的融合蛋白MBP-NS1为包被抗原的间接ELISA检测方法，为AIV灭活疫苗免疫家禽与AIV感染家禽的鉴别诊断奠定了基础。     

日粮添加油脂和出生类型对羔羊肌肉胶原蛋白特性及相关基因表达的影响

旨在研究日粮添加油脂和不同出生类型对羔羊肌肉胶原蛋白特性及相关基因表达的影响。本试验选取64只2月龄断奶双羔(twin born,TW;32只)和三羔(triplet born,TR;32只)湖羊公羔,分别饲喂基础日粮(control, CON)和添加2%大豆油(soybean oil,SBO)的日粮,采用2×2析因试验设计,分为4个处理(TW-CON,TW-SBO,TR-CON,TR-SBO),每个处理16个重复,每个重复1只羊。90 d后屠宰采集羔羊背最长肌,测定其胶原蛋白含量、相关酶活以及肌肉发育相关基因表达水平等。结果表明:1) TW组的Ⅰ型胶原蛋白含量显著高于TR组(P<0.05)。日粮添加油脂有提高可溶性胶原蛋白含量的趋势(0.050.05)。2)基质金属蛋白酶13(MMP-13)含量因日粮中油脂添加而显著下降(P<0.05),而吡啶诺林(PYD)和基质金属蛋白酶抑制剂(TIMP-1)则受日粮油脂添加和出生类型交互作用的显著影响(P<0.05)。3)MYOD1在双羔中表达水平显著高于三羔(P<0.05)。日粮油脂添加显著降低了FN1的相对表达水平(P<0.05),而FGF2和COL3A1的相对表达水平则表现出受日粮油脂添加和出生类型交互作用影响的趋势(0.050.05)。结果提示,双羔肌肉中胶原蛋白含量高于三羔,油脂有促进胶原蛋白含量增加的趋势,且日粮油脂和出生类型共同作用胶原蛋白代谢相关酶含量及基因的表达,进而影响肌肉中胶原蛋白的含量。