SNP的研究进展及其在家畜育种中的应用

单核苷酸多态性(SNP)作为新的遗传标记,已广泛应用于动物遗传育种、基因定位、克隆和遗传多样性等方面的研究。文章对SNP的概念、检测方法进行了详细阐述,并综述了SNP在动物遗传育种和家畜繁殖技术中的应用。     

基于16S rRNA和Cyt b基因序列探讨2种梅童鱼的遗传分化

比较分析了棘头梅童鱼（Collichthys lucidus）和黑鳃梅童鱼（C.niveatus）的16SrRNA和Cytb基因片段序列差异及遗传分化程度。在长度为526bp的16SrRNA和379bp的Cytb基因片段的核苷酸序列中,2种间共检测到44处核苷酸替代。分析结果表明,2个基因片段的鸟嘌呤（G）含量较低,在Cytb蛋白质编码基因第三密码子位点上表现尤为明显。基于16SrRNA和Cytb基因片段分析结果显示,2种间平均遗传距离分别为0．012和0．111。构建的系统树显示2种梅童鱼在这2个基因片段上存在显著的遗传分化。根据Cytb基因2％／百万年的进化速率推断,棘头梅童鱼与黑鳃梅童鱼的分化时间约为550万年,发生于上新世（Pliocene）早期。     

猪血超氧化物歧化酶(SOD)性质的研究

本文对猪血红细胞精制SOD的理化性质进行了研究。研究结果表明精制猪血SOD分子量为31697.86,亚基分子量15848.9,SDS-PAGE为1个条带。DAGE电泳为4个条带。FPLC法测定SOD为单一的吸收峰,达均一纯度。人源、牛源、猪源SOD氨基酸序列比较,猪源与人源的SOD在一级结构中有27个氨基酸不同,占总数的17.6%;牛源与人源的SOD在一级结构中有28个氨基酸不同,占总数的18.3%;仅从SOD氨基酸排列的序列上看,猪源SOD较牛源SOD更接近人源SOD。     

中国近海与日本近海白姑鱼线粒体细胞色素b基因全序列比较分析

对中国近海和日本近海共19尾白姑鱼(Pennahia argentata)线粒体细胞色素b(Cytochrome b,Cytb)基因全序列进行扩增与测定,白姑鱼个体的Cytb基因总长度均为1141bp。序列分析结果显示:Cytb基因全序列中共检测到40处核苷酸替代,全部为同义替换,且主要来自密码子第三位点。19尾白姑鱼个体共定义了14种单倍型。核苷酸组成分析表明:白姑鱼Cytb全序列的鸟嘌呤(G)含量较低,在第三密码子位点上表现得尤为明显。基于邻接法、最大简约法、最大似然法和贝叶斯法构建的系统树结果基本一致,中国近海和日本近海的白姑鱼明显分为2个单系群,两组群间的净遗传距离为0.019,基于Cytb2%/百万年分子钟计算,其分化时间约为95万年,中、日近海白姑鱼分化事件发生于晚更新世(Late Pleistocene)。本研究旨在探讨中、日近海白姑鱼组群的分类地位的遗传学依据,为白姑鱼渔业资源的管理与保护提供基础参考。     

SNP分子标记技术在经济甲壳动物中的应用进展

作为第三代分子标记,单核苷酸多态性标记(SNP)具有丰富度高、密度大、稳定性强、共显性等特点,成为目前虾、蟹等经济甲壳动物中应用最广泛的分子标记技术。本文对经济甲壳动物中SNP技术的发展及其在高密度遗传连锁图谱的构建、种质资源遗传多样性分析、分子辅助育种等方面的应用进展进行了综述,并提出了未来有待加强的研究方向,以期为经济甲壳动物种质资源的开发和利用提供理论指导。     

细菌性鱼病快速诊断技术的进展

简要地归纳了当前国内外学者应用现代生物技术检测细菌性鱼病的手段，主要有：荧光抗体技术、免疫酶技术、单克隆抗体技术、核酸杂交技术、聚合酶链反应（ＰＣＲ）技术、以及电子计算机应用技术等，反映了鱼病学实用诊断技术的进展情况     

Complete nucleotide sequence and variation of mitochondrial DNA from 10 individuals of walleye pollock, Theragra chalcogramma

ABSTRACT:   The complete mitochondrial genome sequence of 10 walleye pollocks, Theragra chalcogramma , from the Japan Sea and Bering Sea was determined. The 16 568–16 571 bp genome contains the same 37 mitochondrial structural genes (two ribosomal RNA, 22 transfer RNA, and 13 protein-coding genes) as found in all other vertebrates analyzed, in an organization identical to that of other bony fish. The major non-coding region had several conserved sequence features. Nucleotide variations of ND1, ND5, and control region were high, and these regions appear to be good candidates for high-resolution markers in population studies.     

Development of a Penaeus monodon-type baculovirus (MBV) DNA probe by polymerase chain reaction and sequence analysis

Abstract. Penaeus monodon -type baculovirus (MBV) was isolated and purified from the hepatopancreases of MBV-infected Penaeus monodon Fabricius. MBV DNA was extracted and used as a template in a polymerase chain reaction (PCR). The primers were chosen from conserved regions of the polyhedrin gene of Autographa californica nuclear polyhedrosis virus (AcNPV). One DNA fragment (674 base pairs) was amplified after PCR. There was a 65% homology between the predicted amino acid sequence of this PCR product with that of the polyhedrin polypeptide of AcNPV. Nucleotide sequence analysis indicated that the amplified DNA is the open reading frame of the MBV polyhedrin gene. This 674 bp DNA fragment was subsequently used as a probe in a dot blot analysis. The probe was able to hybridize with the DNA extracted from the purified MBV and from the MBV-infected P. monodon , but not from the MBV uninfected P. monodon.     

鲫胰岛素样生长因子-Ⅰ cDNA克隆及序列分析

利用One-Step RT-PCR技术,从鲫(Carassius auratus)肝胰脏组织中克隆了胰岛素样生长因子-Ⅰ(IGF-Ⅰ),并进行了序列分析。结果表明,鲫IGF-ⅠcDNA由486个核苷酸组成,编码包括信号肽、成熟肽(B、C、A、D四个域)和E域的161个氨基酸。核苷酸同源性分析揭示,鲫IGF-ⅠcDNA序列与金鱼的同源性最高,为99.18%;与鲮、三角鲂、团头鲂的同源性次之,分别为94.44%、94.65%、94.44%;与鲤、草鱼的同源性相对较低,为86.03%和85.29%。E区域的分析结果表明,鲫IGF-Ⅰ序列为Ea-2亚型。     

饲料中添加外源核苷酸对动物生长的影响

核苷酸作为一种无毒无害无残留的新型饲料添加剂,对动物生长发育和免疫功能等具有重要作用,在动物营养方面有很大的发展前途。近年来国内外学者对核苷酸的研究逐渐增多,本文综述了添加外源核苷酸对动物肠道功能的影响,进而对动物生长的影响。     

常规PCR、实时定量PCR检测传染性皮下及造血器官坏死病毒的效果分析

对虾传染性皮下及造血器官坏死病毒(IHHNV)可感染世界各地养殖对虾,给对虾养殖业造成严重经济损失。本实验首次采用实时定量PCR法对广西地区的84份凡纳滨对虾样品进行检测,同时以常规PCR检测作对照。实时定量PCR检测阳性率为79·8%,常规PCR检测阳性率为40·5%,表明广西地区养殖的凡纳滨对虾IHHNV的感染率较高。将二者检测均呈阳性的30份样品扩增产物进行序列分析测序,测序结果通过DNA STAR软件包进行分析,并通过NCBI Blast与GenBank中的序列进行比对。结果证明,测定的是IHHNV序列。30份样品的IHHNV序列很保守,可以分为4种类型,仅有两个碱基的位置发生变异。实时定量PCR检测IHHNV,快速、灵敏、准确,特异性好,可以作为检测对虾感染病毒的有效方法。     

Molecular cloning of Yersinia ruckeri aroA gene: a useful taxonomic tool

The aroA gene of Yersinia ruckeri , which encodes 5-enolpyruvylshikimate 3-phosphate (EPSP) synthase was cloned by complementation of the aroA mutation in Escherichia coli AB2829 by using pUC18 plasmid as a vector. Nucleotide sequence of the aroA gene revealed an open reading frame of 427 amino acids showing a high degree of homology to other bacterial AroA proteins. A pair of primers with 23 and 20 nucleotides were selected from the 5' and 3' termini, respectively, and formed the basis of a specific polymerase chain reaction (PCR) assay. A 1165-bp deoxyribonucleic acid (DNA) fragment was amplified from all lysed Y. ruckeri strains. An identical size fragment was also amplified from lysed Y. pseudotuberculosis , Y. aldovae , Salmonella enteritidis and E. coli , but not from other enterobacteria. Alu I restriction fragment length polymorphism (RFLP) of the PCR amplified products allowed for differentiation between Y. ruckeri and the other bacteria. Specificity and sensitivity make this PCR assay a useful method for rapid identification and diagnosis of Y. ruckeri infections.     

Nucleotide‐binding and oligomerization domain (NOD)‐like receptors in teleost fish: Current knowledge and future perspectives

Nucleotide‐binding and oligomerization domain (NOD)‐like receptors (NLRs) are a group of intracellular pathogen recognition receptors (PRRs) that play key roles in pathogen recognition and subsequent activation of innate immune signalling pathways. Expressions of several NLR subfamily members, including NOD1, NOD2, NLR‐C3, NLR‐C5 and NLR‐X1 have been reported in many different teleost fish species. These receptors are activated by a variety of ligands, including lipopolysaccharides (LPS), peptidoglycans (PGN) and polyinosinic‐polycytidylic acid [Poly(I:C)]. Synthetic dsRNA and bacterial or viral infections are known to stimulate these receptors both in vitro and in vivo. In this review, we focus on the identification, expression and function of teleost NLRs in response to bacterial or viral pathogens. Additionally, NLR ligand specificity and signalling pathways involved in the recognition of bacterial or viral stimuli are also summarized. This review focuses on current knowledge in this area and provides future perspectives regarding topics in need of additional investigation. Understanding the response of innate immune system to bacterial or viral infections in diverse species could inform the development of more effective therapies and vaccines.     

两种黄盖鲽线粒体DNA部分片段比较分析

比较分析了钝吻黄盖鲽（Pleuronectes yokohama）和尖吻黄盖鲽（P. herzensteini）线粒体基因组COI、Cyt b基因以及D-Loop片段总长度为1373 bp的核苷酸序列,两种间共检测到107处核苷酸替换,蛋白质编码基因上的核苷酸替代主要是第三密码子位点上的同义替换。核苷酸组成分析结果表明：三个目的片段的鸟嘌呤（G）含量普遍较低,在两个蛋白编码基因第三密码子位点上表现得尤为明显。两种黄盖鲽在线粒体基因组不同片段上存在明显的遗传分化,核苷酸替代速率最快的是D-Loop,COI和Cyt b核苷酸替代速率基本一致。建议在进行黄盖鲽属鱼类分子系统发育研究时,应该针对不同研究目的选择合适的分子标记。基于Cyt b基因片段序列的分析结果表明：钝吻黄盖鲽和尖吻黄盖鲽两种的分岐时间约为200万年,两种间的分化事件发生在更新世（Pleistocene）。     

大泷六线鱼6个野生群体遗传多样性的12S rRNA基因分析

本文利用PCR技术扩增了6个野生群体大泷六线鱼线粒体12S r RNA基因的部分序列。在获得的582bp碱基序列中,共检测到314个变异位点,占总位点数的53.95%。34尾个体共定义了16种单倍型,6个群体的单倍型多样性指数为0.40~1.00,核苷酸多样性指数为0.00071~0.10441。中性检验Fu's Fs值为1.19782(P0.01)。分子变异分析表明:Fst=0.88326(P0.01),群体间和群体内变异分别为88.33%和11.67%。不同个体间的遗传距离构建的NJ和MP系统树表明,东港和旅顺群体亲缘关系较近,构成一个分支,其他群体构成另一分支。     

脊尾白虾3个野生群体ITS1序列分析及其亲缘关系分析

对脊尾白虾的莱州湾、海洲湾、象山湾3个野生群体共计62个个体的核糖体RNA转录单元内间隔区ITS1基因片段进行克隆和测序,对序列特点进行分析,并结合GenBank数据库中已有的长臂虾亚科ITS1同源序列虾类进行系统分析。结果显示,脊尾白虾的ITS1序列具有长度多态性,其长度为345~384 bp,62条序列GC的平均含量显著高于AT含量;共检测到79个变异位点,39种单倍型,多态位点比例为21.7%;海洲湾群体遗传多样性指数最高,象山湾群体次之,莱州湾群体最低。在脊尾白虾ITS1序列中发现微卫星序列共有8处,重复序列类型为(GC)n、(AG)n、(GGC)n、(GGA)n、(AT)n、(GA)n,以(GA)n类型最多。AMOVA分析结果显示3个群体间的遗传分化较弱或只有中度分化。另外用MEGA4.0软件中的NJ法构建分子进化树,探讨长臂虾亚科几个种的系统进化关系,系统树显示同种的不同个体、同属的不同种聚在一起,与形态学的分类吻合。     

Koi herpesvirus and carp oedema virus: Infections and coinfections during mortality events of wild common carp in the United States

Koi herpesvirus (KHV; cyprinid herpesvirus‐3) and carp oedema virus (CEV) are important viruses of common and koi carp (Cyprinus carpio); however, the distribution of these viruses in wild common carp in North America is largely unknown. During the summers of 2017 and 2018, 27 mass mortalities of common carp were reported from four states in the USA (Minnesota, Iowa, Pennsylvania and Wisconsin), the majority of which were distributed across eight major watersheds in southern Minnesota. Samples from 22 of these mortality events and from five clinically healthy nearby carp populations were screened for KHV, CEV and SVCV using real‐time polymerase chain reaction (qPCR). KHV was confirmed in 13 mortality events, CEV in two mortality events and coinfections of KHV/CEV in four mortality events. Nucleotide sequence analysis revealed that the KHV and CEV detected here are closely related to European lineages of these viruses. While molecular detection alone cannot conclusively link either virus with disease, the cases described here expand the known range of two important viruses. This is also the first reported detection of KHV and CEV coinfections in wild carp populations.     

Isolation and characterization of bacteria causing mortality in early stage larvae of captive‐bred Japanese eels (Anguilla japonica Temminck & Schlegel)

Bacteria with lethal effect on eel larvae were isolated from moribund captive‐bred eel larvae and their 16S rRNA gene sequences were analysed. Nucleotide sequence of 10 isolates showed highest similarity with Lacinutrix algicola, Crocinitomix catalasitica and Pseudoalteromonas rubra. Age‐dependent changes in the susceptibility of eel larvae were observed in response to challenge by a highly lethal isolate. Compared with 10 and 11 days after hatching (DAH), larval susceptibility to the highly lethal isolate was lower at 18 DAH. We found that the bacterial isolates have lethal effect on the captive‐bred eel larvae, especially at the early developmental stage. These results will be useful to establish appropriate culture practices for eel larvae that will improve the success of mass production of glass eels for aquaculture.