关岭牛MyoD Ⅰ基因在不同组织和时期中的表达分析

为研究MyoDⅠ基因在关岭牛不同组织表达量的差异及在肌肉发育过程中的表达情况,试验采用实时荧光定量PCR技术对关岭牛背最长肌、大腿肌、心脏、肝脏、脂肪、小肠6个组织的MyoDⅠ基因相对表达量进行检测。同时分析关岭牛胎儿、18月龄、30月龄时期背最长肌组织中MyoDⅠ基因的表达规律。结果显示,MyoDⅠ基因在背最长肌中表达量最高,在出生后的关岭牛肌肉组织中也具有较高的表达量,且随着出生时间的推移呈上升趋势。推测可能由于背最长肌是肌肉富集地方,因此基因表达量较高,随着年龄增大,肌肉丰满度增加,基因表达量也随着增加。     

红毛藻中多组分藻胆蛋白及其亚基的分离

采用DEAE-52离子交换柱对红毛藻(Bangia fusco-purpurea Lyngb.)中的藻胆蛋白进行分离，依次洗脱出蓝色、紫色、深红色、红色和浅红色等几种组分的藻胆蛋白；经光谱分析发现蓝色组分为R-藻蓝蛋白，紫色组分为1种三峰型的藻胆蛋白，深红色组分和红色组分为2种不同但又非常相似的藻红蛋白，其中深红色组分可能与某些色素物质结合较为紧密，而最后洗脱出的浅红色组分则为变性的藻红蛋白。尿素洗脱结果表明，蓝色组分的R-藻蓝蛋白是由2种双峰型的亚基构成。本研究旨为揭示藻胆蛋白的能量传递机制提供理论和技术依据。     

东方粘虫中肠V-ATPase H亚基原核表达及纯化

对东方粘虫Mythimna separate(Walker)中肠V-ATPase H亚基基因(VATPH)进行克隆、原核表达及纯化。首先采用RT-PCR技术克隆基因,再构建原核表达载体(pET15b-VATPH)并转入E.coil BL21(DE3),经异丙基硫代-β-D-半乳糖苷(isopropyl-β-D-thiogalactoside,IPTG)诱导表达,用Ni-NTA柱及S-200分子筛纯化,最后进行SDS-PAGE分析。结果表明,pET15b-VATPH可高效表达V-ATPase H亚基,纯化后可获得单一条带且分子质量约为55ku的重组蛋白。该结果为进一步研究V-ATPase H亚基的晶体结构,药物小分子与H亚基大分子的相互作用,尤其是以H亚基为筛选模型创制新农药奠定基础。     

开花麻竹叶绿素荧光参数测定

叶绿素荧光是研究光合作用的有效探针。为了探讨开花对麻竹光合作用的影响,应用PAM-100分别测定了开花与未开化麻竹的叶绿素荧光参数。结果表明,随着光强的升高[0~2 000μmol/（m^2·s）],在开始阶段[0~200μmol/（m^2·s）]麻竹叶片的NPQ、Y（NPQ）、Y（ND）、ETR（Ⅱ）和ETR（Ⅰ）均迅速升高,Y（Ⅱ）和Y（Ⅰ）却迅速下降,之后变化缓慢并趋于平稳,而Y（NA）和Y（NO）几乎一直维持平稳;其中未开花麻竹的NPQ、Y（Ⅰ）和ETR（Ⅰ）均高于开花麻竹,Y（Ⅱ）、Fv/Fm和ETR（Ⅱ）无明显差异。在本实验培养光照下[230μmol/（m2·s）],未开花麻竹的NPQ、Y（Ⅰ）和ETR（Ⅰ）也均高于开花麻竹。由此表明,开花引起麻竹PSⅠ的Y（Ⅰ）和ETR（Ⅰ）以及PSⅡ的NPQ降低,这意味着开花麻竹的光保护能力下降,使得PSⅠ受体侧电子积累,导致光合效率下降。     

猪传染性胃肠炎病毒S基因实时荧光定量RT-PCR检测方法的建立及初步应用简

据猪传染性胃肠炎病毒（transmissible gastroenteritis virus of swine,TGEV）S基因序列设计1对特异性引物,通过对实时荧光定量RT-PCR反应条件的优化,建立了SYBR GreenⅠ实时荧光定量RT-PCR检测TGEV的方法,同时对12份病料进行检测,并与常规RT-PCR进行比较。结果显示,该方法的敏感性达到43.07拷贝/μL,具有良好的特异性和重复性,而常规RT-PCR最低只检测到4.307×10³ 拷贝/μL,敏感性较低。本试验建立的检测TGEV S基因的SYBR GreenⅠ实时荧光定量RT-PCR方法为传染性胃肠炎的鉴别诊断及TGEV的分离鉴定奠定了技术基础。     

Cloning and Sequence Analysis of Major Histocompatibility Complex Ⅰ Gene from Goose F1 of Fast-growth Lines

According to the multiple alignments identified major histocompatibility complex Ⅰ (MHC Ⅰ) gene conserved sequence registered in GenBank from the family ducks (Anatidae) anser waterfowl, a pairs of specific primers for the fragments of MHCⅠgene of goose F1 from fast-growth lines were designed and synthesized by Primer Premier 5.0. Using the genome DNA of goose F1 from fast-growth lines, the target gene fragment was obtained by PCR. To conduct sequencing of the fragments of MHCⅠgene of goose F1 from fast-growth lines and make sequence alignment and analysis of protein structure and function by bioinformatics, and research the characteristics of MHCⅠgene of goose F1 and the physicochemical properties of the protein. Bioinformatics was analyzed the nucleic acid data, deduced amino acid sequence and phylogenetic trees. The result of sequence analysis showed that the fragments of MHCⅠgene of goose F1 from fast-growth lines was 1036 bp in length, which coded 96 amino acids polyprotein. The homology were 93% and 83% with MHC Ⅰ gene and coding sequence of Wulong goose in NCBI respectively. There were 72 different bases sequence and 16 amino acids change. There also was higher homology with other poultry, and existed genetic relationship of Siji goose > chickens > ducks.The homology segment sequences corresponding to the fragments of MHCⅠ gene of goose F1 coded 96 amino acids protein, which molecular weight, PI, positively or negatively charged amino acid, estimated half-life, instability index, aliphatic index and average hydrophobicity were 11.342 ku, 5.32, 14, 17, 2.8 h, 34.92, 42.81, -1.066, respectively, and appeared 9 B cell epitopes, but contained no signal peptide. These results indicated that the protein for hydrophilic non-secreted proteins, had the high immunogenicity. In addition, The protein structure study indicated that alpha-helix, beta-sheet, beta-turn and random coil were 31.25%,16.67%, 14.58% and 37.50%, respectively. There existed amino terminal domain and carboxyl terminal domain in the tertiary structure. Therefore, MHC gene had significant difference between species and populations of individuals by the pathogen pressure in environment, and there were the interaction between polymorphism of MHC molecules and the diversity of antigenic peptide. MHC determined the differences of individual susceptibility to disease, and could be treated as a candidate gene for disease resistance.     

羊绒与羊毛的PCR-RFLP识别方法

羊绒价格昂贵,在销售中时常出现掺假现象,最常见的是用羊毛,因此精确识别羊绒和羊毛纤维的方法非常重要。本试验采用聚合酶链式反应-限制性片段长度多态（PCR-RFLP）的技术方法,利用提取的羊绒和羊毛线粒体基因组,根据细胞色素b（Cytochrome b）基因已知的DNA序列设计引物,通过内切酶SspⅠ进行酶切,得到不同的片段长度,根据酶切图谱鉴别羊绒和羊毛样品,用来进行羊绒掺假试验的定性检测。     

Isolation of a novel hepatic CYP2-related protein from channel catfish, Ictalurus punctatus

Channel catfish (Ictalurus punctatus) have been previously shown to express two major cytochrome P450 (CYP) protein bands that are cross-reactive with anti-CYP2K1 (rainbow trout, Oncorhynchus mykiss) antibodies on Western blots. These proteins appear to be the major constitutive CYPs in channel catfish and show distinct sex- and age-specific variations in expression. Because I. punctatus is an important agricultural and ecological commodity, and because it displays a high degree of resistance to the toxic effects of many pesticides, the molecular and catalytic characteristics of its biotransformation systems are of interest to those in areas of environmental science and aquaculture research. Using a chromatographic method similar to that employed in the purification of other fish CYP2 enzymes, a single CYP2-related protein (CM-HA3) was isolated from channel catfish hepatic microsomes. The isolated protein displays a relative molecular mass of approximately 47 kDa, and a CO-reduced difference spectrum _max of 449.6 nm. The sequence of 15 residues at the amino-terminal of CM-HA3 is 27% identical to both CYP2K1 and CYP2M1 isoforms of rainbow trout. Correlational analysis was employed to characterize potential substrates for this isoform, but no significant relationship was observed with E₂ hydroxylation, testosterone hydroxylation, or 7-ethoxycoumarin O-deethylase activities. These data indicate that CM-HA3 is a CYP2 family protein, with as yet uncharacterized substrate specificities.     

江苏省小麦品种的谷蛋白亚基组成分析

为了解江苏省小麦品种的谷蛋白亚基组成现状,为品质育种提供理论指导,选用77份来自江苏省淮南和淮北主要小麦育种单位的育成品种或高代品系,利用聚丙烯酰胺凝胶电泳法(SDS-PAGE)对Glu-1和Glu-3位点进行了鉴定。结果表明,参试品种(系)Glu-1和Glu-3位点的高分子量麦谷蛋白亚基(HMW-GS)和低分子量麦谷蛋白亚基(LMW-GS)变异丰富。共检测到12个HMW-GS等位变异,其中Glu-A1位点有3个,Glu-B1位点有5个,Glu-D1位点有4个。LMW-GS的Glu-A3和Glu-B3位点则分别检测到5个和6个等位变异。淮北小麦的HMW-GS多态性高于淮南小麦。Glu-1和Glu-3位点等位变异的分布频率差异较大,且淮北和淮南表现不同,优质亚基比例较低。Glu-1位点主要亚基类型为0(46.8%)、1(49.4%)、7+8(63.6%)和2+12(63.6%),Glu-A3位点主要亚基类型为a(23.4%)、c(57.1%)和d(15.6%),Glu-B3位点主要亚基类型为b(22.1%)、f(49.4%)和j(1B/1R易位)(16.9%)。淮北小麦的高分子量谷蛋白优质亚基及亚基组合较少,Glu-B3j分布频率高(54.5%),亚基质量有待提高;淮南小麦的优质亚基及组成亦较少,需根据中筋和弱筋小麦育种目标并结合蛋白质含量和亚基选择进行改良。     

动物布鲁氏菌病疫苗的来历及亚单位疫苗的研究概况

布鲁氏菌病是一种严重危害人和动物健康的人畜共患病,人群感染主要来源于感染的动物及被污染的畜产品,目前疫苗免疫仍是控制动物布鲁氏菌病的主要措施。现有国际参考疫苗主要有牛种疫苗株S19和羊种疫苗株Rev.1,这些疫苗为动物群布鲁氏菌病控制提供重要保障的同时也存在关键的技术瓶颈问题,如安全性差,接种动物出现流产;血清抗体体内持续时间较长,干扰常规诊断等。随着布鲁氏菌基因组测序工作的相继完成及DNA重组技术的发展,新型布鲁氏菌病疫苗的开发成为研究的热点。亚单位疫苗本着其较高的安全性和有效性,在动物群布鲁氏菌病控制过程中具有较大的应用前景。笔者综述了布鲁氏菌病疫苗的发展史,并对布鲁氏菌病亚单位疫苗的研究进展进行概述。