杜氏盐藻基因DsSP的克隆、分析和原核表达(英文)

[目的]克隆盐藻淀粉磷酸化酶基因,并初步分析其基本性质和表达蛋白。[方法]采用RT-PCR和RACE的方法克隆基因;生物信息学系统分析其性质;构建原核表达载PGS21a-DsSP转化于E.coli BL21表达蛋白,采用GST-SefinoseTMKit和Western Blot分别纯化、检测该融合蛋白。[结果]成功克隆出1种淀粉磷酸化酶(GenBank accession No.KF061044)命名为DsSP;分析得到了其基本性质并预测了该蛋白的亚细胞定位、二级结构和三级结构;该融合蛋白在上清和包涵体中均存在,且对上清蛋白成功纯化;Western Blot检测表明其在E.coli.BL21中成功表达。[结论]为进一步阐明DsSP的功能及作用机制奠定了理论基础。     

输卵管炎种鸭大肠杆菌的分离鉴定及生物学特性分析

为分析表现输卵管炎种鸭体内分离的大肠杆菌的致病性、药敏试验、血清型及毒力基因携带情况,无菌采集表现输卵管炎的种鸭肝脏,进行大肠杆菌的分离鉴定,经易感鸭皮下注射,测定其致病性;纸片法进行药敏试验;凝集试验确定血清型;PCR法检测毒力基因.结果共分离鉴定大肠杆菌16株,其中强毒菌株11株.70％以上菌株表现敏感的药物有丁胺卡那霉素、氟苯尼考、头孢唑啉、左氧氟沙星和壮观霉素,70％以上菌株表现耐受的药物有复方新诺明、卡那霉素、庆大霉素、恩诺沙星和氟哌酸.16株大肠杆菌分属于4个血清型,078占56.3％,为主要血清型.毒力基因irp2、yuA、imC和pa pC的检出率分别为68.8％、87.5％、75％和100％.总结说明,从表现输卵管炎的种鸭体内共分离到16株大肠杆菌,078是主要的血清型,首选壮观霉素和氟苯尼考等敏感药物进行治疗.     

胰腺炎型鸭1型甲肝病毒结构蛋白VP1基因的克隆和表达

通过RT-PCR方法扩增出MPZJ1206株胰腺炎型鸭1型甲肝病毒结构蛋白VP1基因,将其与原核表达载体pGEX-6P-1连接获得重组表达质粒pGEX-VP1,进行条件优化诱导表达,将表达的重组蛋白经SDS-PAGE分析。结果表明,分子量约为52 kDa的重组目的蛋白得到表达。该研究为开发胰腺炎型鸭1型甲肝病毒诊断方法和研究VP1蛋白功能奠定基础。     

基于线粒体COI基因的17种菱蜡蝉亚科昆虫DNA条形码研究（半翅目：蜡蝉总科：菱蜡蝉科）

通过对菱蜡蝉科Cixiidae菱蜡蝉亚科Cixiinae 9属17种昆虫的mt DNA COI基因序列进行研究,探讨DNA条形码在菱蜡蝉亚科昆虫中快速识别和准确鉴定的可行性.采用MEGA 5对序列进行比对和遗传距离分析,基于COI基因序列构建ML、MP、NJ、ME系统发育树.结果显示：属间平均遗传距离为0.133,介于0.102 ～0.147之间;属内种间平均遗传距离为0.047,介于0.025～ 0.079之间;地理种群间平均遗传距离为0.039;介于0.024～0.064之间.系统发育树显示：同属物种聚为一小支,分支置信度高达97％ ～ 100％;同一地理类群聚为一支,分支置信度高达98％ ～100％.结果表明应用基于COI基因片段的DNA条形码对菱蜡蝉亚科昆虫分类鉴定是可行的.     

鹰嘴豆LHY基因cDNA克隆及生物信息学分析

从鹰嘴豆中克隆生物节律钟LHY基因的cDNA全长序列,进行序列信息学分析。通过同源克隆策略,利用RT-PCR技术获得核心片段,结合5＇-RACE和3＇-RACE技术,克隆得到鹰嘴豆生物节律钟基因LHY的cDNA全长序列,其核苷酸序列长度为3 061 bp,包括2 220 bp的完整开放阅读框（ORF）,编码739个氨基酸。验证后命名为CarLHY基因,获得基因登录号为KJ558378。生物信息学研究表明CarLHY基因cDNA序列与其他植物LHY基因具有较高的相似性;预测CarLHY蛋白不具有跨膜结构;为转录因子,定位于细胞核中;不具备信号肽。对CarLHY蛋白功能结构域预测表明,蛋白质核心结构存在符合转录因子与DNA结合的常见功能域HTH。蛋白系统进化树显示,与大豆分子进化距离最近,其次是黑杨、拟南芥。     

荔波瑶山鸡催乳素基因5′-调控区24-bp插入／缺失变异与早期产蛋性状的关系

为提高贵州地方鸡种荔波瑶山鸡的产蛋性能,以催乳素基因 PRL 为候选基因之一,利用 PCR 方法直接检测催乳素基因 PRL 的5′-调控区24-bp插入／缺失（－377～－354bp）变异情况,并分析其与产蛋性能的关系。结果表明,荔波瑶山鸡该位点83．8％为缺失型,无插入纯合子;不同基因型（PRL＋／＋、＋／－和－／－型）对荔波瑶山鸡早期产蛋性能均无显著影响（P＞0．05）。     

Nitrogen Deficiency Limited the Improvement of Photosynthesis in Maize by Elevated CO2 Under Drought

Global environmental change affects plant physiological and ecosystem processes. The interaction of elevated CO2, drought and nitrogen （N） deficiency result in complex responses of C4 species photosynthetic process that challenge our current understanding. An experiment of maize （Zea mays L.） involving CO2 concentrations （380 or 750 μmol mol1, climate chamber）, osmotic stresses （10% PEG-6000, -0.32 MPa） and nitrogen constraints （N deficiency treated since the 144th drought hour） was carried out to investigate its photosynthesis capacity and leaf nitrogen use efficiency. Elevated CO2 could alleviate drought-induced photosynthetic limitation through increasing capacity of PEPC carboxylation （Vp~,x） and decreasing stomatal limitations （SL）. The N deficiency exacerbated drought-induced photosynthesis limitations in ambient CO2. Elevated CO2 partially alleviated the limitation induced by drought and N deficiency through improving the capacity of Rubisco carboxylation （Vmax） and decreasing SL. Plants with N deficiency transported more N to their leaves at elevated CO2, leading to a high photosynthetic nitrogen-use efficiency but low whole-plant nitrogen-use efficiency. The stress mitigation by elevated CO2 under N deficiency conditions was not enough to improving plant N use efficiency and biomass accumulation. The study demonstrated that elevated CO2 could alleviate drought-induced photosynthesis limitation, but the alleviation varied with N supplies.     

Molecular Mapping of a Stripe Rust Resistance Gene YrH9020a Transferred from Psathyrostachys huashanica Keng on Wheat Chromosome 6D

Stripe rust （yellow rust）, caused by Puccinia striiformis f. sp. tritici （Pst）, is one of the most devastating diseases of wheat throughout the world. H9020-1-6-8-3 is a translocation line originally developed from interspeciifc hybridization between wheat line 7182 and Psathyrostachys huashanica Keng and is resistant to most Pst races in China. To identify the resistance gene（s） in the translocation line, H9020-1-6-8-3 was crossed with susceptible cultivar Mingxian 169, and seedlings of the parents, F1, F2, F3, and BC1 generations were tested with prevalent Chinese Pst race CYR32 under controlled greenhouse conditions. The results indicated that there is a single dominant gene, temporarily designated as YrH9020a, conferring resistance to CYR32. The resistance gene was mapped by the F2 population from Mingxian 169/H9020-1-6-8-3. It was linked to six microsatellite markers, including Xbarc196, Xbarc202, Xbarc96, Xgpw4372, Xbarc21, and Xgdm141, lfanked by Xbarc96 and Xbarc202 with at 4.5 and 8.3 cM, respectively. Based on the chromosomal locations of these markers and the test of Chinese Spring （CS） nullitetrasomic and ditelosomic lines, the gene was assigned to chromosome 6D. According to the origin and the chromosomal location, YrH9020a might be a new resistance gene to stripe rust. The lfanking markers linked to YrH9020a could be useful for marker-assisted selection in breeding programs.     

植物基因打靶的研究与应用

本文分析了影响基因打靶效率的因素,并指出提高同源重组利用率和在靶基因位点处引入DNA双键断裂可以提高基因打靶效率,总结了近年来这两方面尝试在植物中的研究进展,并与一些其他物种中的研究作出比较.     

鸭ApoVLDL-Ⅱ基因第1内含子多态性及其遗传效应

以褐色菜鸭为材料,对其ApoVLDL-Ⅱ基因第1内含子进行PCR-SSCP检测,分析其多态性及遗传效应.结果表明,在ApoVLDL-Ⅱ基因第1内含子检测到多态性,存在BB、Bb和bb 3种基因型,序列比对发现存在T374C、C379T和T390G 3个单核苷酸多态位点,该位点属中度多态,并处于Hardy-Weinberg平衡状态(P>0.05).关联分析显示,bb基因型个体的蛋重极显著高于BB型(P<0.01),bb基因型个体的蛋白重显著高于BB型(P<0.05).推测该片段的多态位点可能对褐色菜鸭蛋重和蛋白重有一定的影响.