肥胖基因研究进展

与肥胖有关的基因已发现很多，但ob基因是研究的重点，其表达产物Leptin的作用机制已有较深入的了解。本文主要介绍了ob基因及其产物Leptin的作用机制、生物学效应、Leptin抵抗现象，以及其它与肥胖有关的基因和蛋白。     

口蹄疫病毒VP1基因的原核可溶性表达

针对重组原核表达载体表达目的蛋白以包涵体形式存在的问题,作者对影响外源蛋白表达的IPTG的浓度、温度、时间等因素均进行了探索,以观察VP1蛋白的可溶性情况。通过PCR技术将VP1基因克隆至pGEM-T载体中,然后将pET-28a(+)和pGEM-T载体分别进行双酶切,构建重组的pET-28a-VP1载体,将重组载体转化至BL21中,诱导后经SDS-PAGE检测可见约29 ku的目的蛋白条带。插入的外源目的蛋白主要以包涵体的形式存在,上清中的可溶性蛋白甚微。试验结果表明,这可能与蛋白本身的氨基酸组成有重要关联。     

Effects of active immunization against myostatin on carcass quality and expression of the myostatin gene in pigs

The study was conducted to investigate the effects of active immunization against myostatin on the titer of myostatin antibody, carcass evaluation, activity of creatine kinase and the expression of the myostatin gene in pigs. Eighteen pigs were allotted into three groups (six pigs per group), and pigs in treatment 1, 2 and 3 were immunized with physiological saline, 1 mg or 4 mg myostatin per pig, respectively. Six pigs were killed by electrical stunning followed by exsanguination at BW of 100 kg. The results indicated that the titer of myostatin antibody was increased in treated groups compared to the control group on day 42 ( P  < 0.01) and d 84 ( P  < 0.01). The carcass lean percentage was significantly increased in the treatment groups compared to the control group ( P  < 0.01), and intramuscular fat was significantly decreased in the 4 mg group compared to the control group ( P  < 0.05). The muscle creatine kinase activity of pigs treated with 1 mg and 4 mg myostatin was lower than the control group. The immunization of myostatin signofocantly decreased the myostatin gene expression levels in muscle. It was concluded that optimal active immunization against myostatin could increase the content of myostatin antibody, suppress the activity of creatine kinase and the expression of myostatin gene, and therefore improve the carcass lean percentage for pigs.     

日粮营养对动物脂肪组织基因表达的影响

脂肪组织是一个强大的分泌器官,它能分泌许多与能量、脂肪代谢相关的酶、激素等.调控能量代谢和脂肪组织生长.本文综述了日粮对动物脂肪组织中基因表达作用的研究状况.并阐述了营养成分对这些基因上下游表达的作用机制.     

福建省羊传染性脓疱病毒F1L、B2L和VIR基因的遗传变异分析

为探明2014年-2015年在福建省流行的羊传染性脓疱病毒(ORFV)遗传变异情况,对10株ORFV流行毒株的F1L、B2L和VIR基因进行克隆、测序及分析。结果表明,10株ORFV F1L基因之间的核苷酸序列同源性为97.6%~100%,与国内株的核苷酸序列同源性为96.8%~99.7%,与NZ2参考株的核苷酸序列同源性为96.3%~97.1%;同NZ2参考株比较,FJ-YT2014缺失2个氨基酸;10株ORFV B2L基因之间核苷酸序列同源性为97.5%~99.9%,与国内株的核苷酸序列同源性为96.7%~99.5%,与NZ2参考株的核苷酸序列同源性为96.7%~97.7%;10株ORFV VIR基因之间的核苷酸序列同源性为95.8%~99.5%,与国内株的核苷酸序列同源性为94.6%~99.6%,与NZ2参考株的核苷酸序列同源性为94.6%~96.4%。基于基因核苷酸序列的遗传进化分析表明,10株ORFV F1L基因与福建省分离株、山西株和新疆株亲缘关系较近;10株ORFV B2L基因与新疆、山西、德国毒株亲缘关系较近;10株ORFV VIR基因与台湾、新疆株亲缘关系较近。结果提示,当前福建省流行的ORFV F1L、B2L和VIR基因尚未出现明显变异,但是其F1L、B2L和VIR基因核苷酸序列之间普遍存在异质性。     

禽流感病毒H9N2亚型三重PCR检测方法的建立

为建立简便快速检测禽流感病毒(avian influenza virus,AIV)并同时区分出H9、N2亚型的方法,本试验根据基因库中H9亚型AIV的HA基因、N2亚型AIV的NA基因及AIV的M基因序列,分别设计了3对针对这3种基因保守序列的引物,建立了AIV H9N2亚型的三重PCR检测方法。应用该方法对H9N2亚型AIV模板进行PCR扩增,可得到3条与试验设计相符的目的条带,分别为313 bp (HA基因)、451 bp (NA基因)和667 bp(M基因);对非H9亚型的N2亚型AIV模板进行扩增,出现2条特异性扩增条带,即451 bp (NA基因)和667 bp(M基因);对非H9、N2亚型AIV模板进行扩增则只出现一条目的条带,即667 bp(M基因);对其他禽呼吸道病原体进行PCR扩增,结果均为阴性。敏感性试验结果显示此三重PCR方法最低检出限为10^-2 ng/μL。应用所建立的三重PCR方法对120份临床病料进行检测的结果与病毒分离鉴定结果一致。各项试验结果均表明,该方法对于禽流感病毒尤其是H9、N2亚型禽流感病毒的检测具有快捷、特异、灵敏的特点。     

Identification and quantification of benzimidazole resistance polymorphisms in Haemonchus contortus isolated in Northeastern Brazil

Haemonchus contortus is the most prevalent nematode in Brazil. The objective of this study was to select 6 populations of H. contortus of known or suspected benzimidazole resistance status and characterize these using quantitative real-time polymerase chain reaction (qPCR) for single nucleotide polymorphisms (SNPs) F200Y, F167Y and E198A in the β-tubulin isotype 1 gene. qPCR was performed using DNA from a pool of 10 adult male H. contortus from a single animal per farm. Faecal egg count reduction test (FECRT) and egg hatch test (EHT) were used to determine the resistance status. Samples were obtained from 6 farms located in 5 counties in the Ceará State: Tauá, Boa Viagem, Quixadá, Santa Quitéria and Solonópole. The inbred-susceptible-Edinburgh (ISE) isolate was used as reference for comparative purposes in the qPCR. Benzimidazole resistance was detected by FECRT on all farms with efficacy values ranging from 0 to 51%. EC50 values as determined by EHT were all above 1.49 μg/ml. High frequencies of the resistant SNPs F200Y and F167Y alleles were detected but no resistance was detected at SNP E198A. Our results suggest that the SNPs F167Y and F200Y are both important for benzimidazole resistance in the studied populations.     

水貂TYR基因T138A位点多态性及其与毛色性状的关联分析

试验旨在检测水貂酪氨酸酶（tyrosinase,TYR）基因T138A位点的多态性,并分析其与水貂毛色表型的相关性。提取5种被毛色型430只水貂血液基因组DNA,采用PCR-RFLP技术,对TYR基因T138A位点进行多态性检测,统计等位基因频率与基因型频率,通过卡方（χ²）独立性检验分析该位点多态性与水貂毛色性状的相关性。结果表明,T138A位点存在2个等位基因T和A,形成TT、TA和AA 3种基因型,AA基因型在吉林白水貂群体中为优势基因型（0.9069）,而TT基因型为金州黑水貂、珍珠水貂、咖啡水貂和银蓝水貂群体的优势基因型,其中在金州黑水貂群体中基因型频率最高（1.0000）。关联分析表明,TYR基因T138A位点的多态性与毛色性状呈极显著相关（P<0.0001）。表明TYR基因T138A位点可能是影响水貂毛色的主控位点或与调控白色被毛表型主控位点连锁的分子标记。     

应用DPO-PCR方法特异性检测志贺氏菌

In this study, a dual-priming oligonucleotide (DPO)-based PCR method for specific detection of Shigella was established using ipaH gene of Shigella as the target gene. The results showed that detection sensitivity of the DPO-PCR method was 1.65×10² CFU/mL. Compared with conventional PCR primers, the DPO primers were easily designed, which simplified the design procedure of PCR primers. DPO primers were not sensitive to annealing temperature, which could efficiently amplify the target gene in the annealing temperature range from 50 to 70 ℃. Moreover, due to the special structure of DPO primers, it had a higher specificity than conventional PCR primers, and none nonspecific amplifications were produced in reaction. In the practical application, tests on 133 samples including frozen/fresh meat, fruits and vegetables, fresh milk, eggs and cooked food by the DPO-PCR method showed that 15 samples were Shigella positive, which were in accordance with the testing results using GB method (GB/T 4789.5-2012), showing good practicality and reliability. The DPO-PCR method provided a new tool for fast and accurate detection of Shigella.