播娘蒿甜菜碱醛脱氢酶基因的克隆和表达分析

采用RT-PCR技术克隆了播娘蒿的甜菜碱醛脱氢酶基因全长cDNA序列(DsBADH)。DsBADHcD-NA序列全长1653bp,其中开放阅读框长1503bp,编码一个由501个氨基酸残基组成的蛋白质,推测的蛋白质相对分子质量为54kD,pI为5.5。序列比对结果表明DsBADH与其它物种的BADHs无论在核酸水平还是在蛋白质水平上都表现较高的同源性,表明BADH基因家族具有较高的保守性。DsBADH基因的氨基酸序列在进化上与同属十字花科植物BADH基因距离较近。蛋白质序列存在一个编码十肽的高度保守序列,该结构在醛脱氢酶中是高度保守的,这些残基可能包含NAD 结合位点及酶催化位点,而且含有与酶功能有关的醛脱氢酶高度保守的氨基酸残基Cys,表明DsBADH可编码活性蛋白。N端存在信号肽,初步将该酶定位于叶绿体。半定量RT-PCR分析表明,DsBADH在根、茎、叶以及角果中均表达,但在角果中的表达显著高于其它组织,而且表明DsBADH受盐诱导正调节表达。     

小麦高分子量谷蛋白亚基1Dx5基因在大肠杆菌中的表达

为了给小麦优质亚基研究奠定基础,将高分子量谷蛋白亚基1Dx5基因的核苷酸序列与载体pRSET A重组,将构建好的质粒转化大肠杆菌BL21(DE3)pLysS菌株,通过IPTG使其得到了成功表达.大肠杆菌中表达的1Dx5亚基在SDS-PAGE上与小麦品种钱尼中的1Dx5亚基具有相似的迁移率.用Western blot技术成功检测到了目的基因产物.通过改变IPTG浓度、诱导时间、培养基成分及初始菌液浓度研究了最适表达条件.结果表明,最适表达条件为:LB培养基,菌液初始浓度(OD600)0.4～0.6,IPTG浓度0.4mmol/L,诱导时间为3～5 h.     

Culture of Macrobrachium rosenbergii (De Man 1879) in experimental cages in a freshwater eutrophic lake at different stocking densities

Macrobrachium rosenbergii (de Man 1879) juveniles (0.4 g) were cultured in experimental cages (L × W × H: 2.5 × 1 × 1 m) in Laguna de Bay, the largest lake in the Philippines. The following stocking densities at four replicates each were used: 15, 30, 60 and 90 prawns m⁻² of cage bottom. The mean sizes at harvest after 5 months of culture ranged from 14.3 g for the highest stocking density to 26.3 g for the lowest. The mean size at harvest, daily growth rate and size class distribution were significantly influenced by stocking density, with those at the lowest stocking density showing significantly better growth and overall proportion of larger prawns. Heterogeneous individual growth (HIG) was fairly evident in all treatments. The percentage of blue‐clawed males was not influenced by treatment but the mean weight was significantly higher in the lower stocking densities. Both the percentage and mean weight of berried females were significantly higher in the lowest stocking density. Survival was the highest in the lower stocking densities (55.3%, 54.0%, 52.7% and 36.9% for 15, 30, 60 and 90 prawns m⁻² respectively). Feed conversion ratio (FCR) improved with decreasing stocking density, ranging from 2.1 to 3. As expected, yield per cropping increased with stocking density and ranged from 450 to 1089 g m⁻² yr⁻¹ of actual cage area. Production values obtained in the cage cultured M. rosenbergii were comparable to or even higher than those reported from pond culture, given that the stocking densities used in this study were generally higher than in ponds. The results show that the farming of M. rosenbergii in cages in lakes is a viable alternative to pond culture and has the potential of improve aquaculture production in lakeshore fish farming communities.     

水稻MYB-MYC基因家族的全基因组鉴定、系统进化和表达模式分析

为了挖掘水稻MYB-MYC基因的功能,通过生物信息学手段对水稻MYB-MYC基因家族进行基因组水平的鉴定,并对其系统进化及表达模式进行分析。本研究在水稻中鉴定到17个MYB-MYC基因,根据系统进化树将它们划分为2个亚家族,不同亚组具有特异的保守基序和基因结构。通过对水稻MYB-MYC基因复制事件的分析发现水稻MYB-MYC基因共产生6个基因复制。此外,转录组数据分析发现水稻MYB-MYC基因在不同组织都存在表达且响应不同非生物胁迫。本试验为水稻MYB-MYC基因功能的研究提供了初步的理论基础。     

芒果MinSPS1基因的克隆及表达载体的构建

蔗糖磷酸合成酶(sucrose phosphate synthase, SPS)既是调控蔗糖合成的关键酶又是限速酶,为了研究其在芒果果实中蔗糖合成的作用机理,本研究从芒果果肉中克隆得到一个与蔗糖合成相关的基因,将其命名为MinSPS1,其cDNA全长序列为3 344 bp,开放阅读框为3 168 bp,编码1 055个氨基酸,蛋白质分子量为118.13 k D,等电点为6.08,对MinSPS1基因编码的蛋白进行系统发育分析,发现其与柑橘具有较近的亲缘关系。采用qRT-PCR对该基因在后熟处理的贵妃芒果肉中的表达量进行分析,结果显示在完熟期贵妃芒果肉中表达量较高,而在青熟期表达量较低。本研究为深入了解芒果蔗糖合成的分子机理,进一步构建了p CAMBIA1301-MiSPS1过表达载体,为MinSPS1的功能鉴定提供理论依据。     

绵山天幕毛虫性信息素通讯系统扫描电镜观察

通过对绵山天幕毛虫性信息素释放系统——雌蛾性腺和性信息素接受系统——雄蛾触角进行扫描电镜观察,发现绵山天幕毛虫性信息素分泌腺位于腹尖末端8~9节之间,是一个可外翻的环状结构;性信息素感受器主要为毛形感器。绵山天幕毛虫性信息素通讯系统的研究为绵山天幕毛虫性信息素的快速提取、分析、合成提供了依据。     

Milk Protein Polymorphism in Kangayam Cattle

This paper reports the milk protein polymorphism, the allele frequencies of variants and the possible linkages among various combinations of milk protein phenotypes in the Kangayam cattle of south India. Milk samples from 156 Kangayam cows were typed by starch gel and polyacrylamide gel electrophoresis for caseins and whey proteins, respectively. All the four milk protein components studied, _s1-casein, -casein, -lactoglobulin and -lactalbumin, exhibited polymorphism with high allele frequencies of 0.9231±0.0151 for _s1-casein C, 0.9263±0.0148 for -casein A, 0.9135±0.0159 for -lactoglobulin B and a relatively high frequency of 0.6218±0.0275 for -lactalbumin A. The mean heterozygosity estimated over all the four milk protein loci was 0.2420. Genetic equilibrium was observed among all the loci studied, except -lactalbumin. Linkage analysis confirmed the non-independence between _s1- and -caseins and between caseins and -lactalbumin phenotypes.     

Dengue hemorrhagic fever with special emphasis on immunopathogenesis

Dengue virus infections are a serious cause of morbidity and mortality in most tropical and subtropical areas of the world; Southeast and South Asia, Central and South America, and the Caribbean. Dengue virus infection can be asymptomatic or causes two forms of illness, dengue fever (DF) and dengue hemorrhagic fever (DHF), which is the severe form of dengue illness and often fatal. Pathogenesis of DHF has been analyzed, and two mechanisms are considered to be responsible. These include dengue serotype cross-reactive immune responses and virulence of the virus. The immunopathological mechanisms include a complex series of immune responses. Rapid increase in the levels of cytokines, especially TNF-, and chemical mediators play a key role in inducing unique clinical manifestations of DHF such as plasma leakage, shock, and hemorrhagic manifestations. It is understood that the process is initiated by infection with a virulent dengue virus, often in the presence of antibodies that enhance dengue virus infection in secondary infection, and then triggered by rapidly elevated cytokines and chemical mediators that were produced by intense immune activation. However, complete understanding of the entire pathological mechanism is far from complete, and further studies are still needed.     

3种木质素降解菌的筛选及对构树降解的研究

[目的]筛选高效降解木质素的菌种。[方法]通过马铃薯琼脂平板培养、马铃薯液体摇瓶培养和变色反应,对采自吉文山枯木中的木质素降解菌进行了筛选,并对其降解构树的卡伯值和纤维素进行了测定。[结果]共筛选出3种对木质素降解能力较强的菌种,分别为JWS-1、JWS-2和向日葵菌核病菌。[结论]为进一步研究生物制浆奠定了基础。     

鸡贫血病毒衣壳蛋白基因的原核表达

[目的]为VP1蛋白的免疫学研究和鸡贫血病毒基因工程疫苗的研制奠定基础。[方法]将1个新克隆的鸡贫血病毒vpl基因（AF448446）在大肠杆菌DE3中进行表达,并对该蛋白进行纯化。[结果]结果表明：已成功构建了表达载体pET30（b^＋）-vp1;VP1蛋白已成功诱导表达并得以纯化;获得的VP1蛋白序列与已发表的VPl蛋白序列存在差异。[结论]试验克隆的vp1基因大小为1347bp,编码449个氨基酸的蛋白。