麦洼牦牛血红蛋白α链基因的克隆测序

从麦洼牦牛脊髓中提取总RNA，利用RT-PCR扩增血红蛋白α链基因，获得了532bp的cDNA，连接到PMD-l8载体上，然后转化TOP-10感受态细胞进行克隆测序。结果显示，牦牛血红蛋白α链基因与普通牛相比存在2个碱基差异，并导致血红蛋白α链71位的1个氨基酸改变。此外，还用电泳方法研究了血红蛋白在新生犊牦牛中的表达。     

牛病毒性腹泻病毒HB-DCZ株NS2-3区基因克隆及序列分析

【目的】对牛病毒性腹泻病毒HB-DCZ株NS2-3区基因进行克隆及序列分析,为阐明本病致病机理,更好的控制BVDV感染、流行提供理论依据。【方法】以牛病毒性腹泻病毒河北分离株(HB-DCZ)基因组RNA为模板,扩增BVDVHB-DCZ株基因组NS2-3片段cDNA。将PCR产物克隆后进行PCR及双酶切鉴定。克隆产物进行序列测定及分析。【结果】HB-DCZcDNA体外扩增获得特异性条带,大小约为665bp,表明该分离毒株基因组中无插入序列。PCR产物克隆,提取重组质粒,扩增获得特异性的DNA条带,长度约为665bp,用EcoRⅠ和HindⅢ双酶切,获得两条DNA片段,长度分别为2600bp和702bp左右,与理论值相符。序列测定结果表明,克隆得到的NS2-3重要区扩增片段共665核苷酸,NS2-3重要区的蛋白质由208个氨基酸残基组成。HB-DCZ毒株NS2-3重要区核苷酸序列与已公开发表的BVDV其他毒株相比的同源性依次为184株99.1%,ZM195株97.4%,Osloss株92.3%,C_(24)V株77%,NADL株76.4%。HB-DCZ在NS2-3扩增区域中既没有外源序列的插入,也没有基因重组、重排或缺失,但存在某些核苷酸的替换。【结论】HB-DCZ与Osloss株、国内的184株、ZM195株遗传距离较近,与C_(24)V株、NADL株遗传距离较远。HB-DCZ株应为BVDVIb基因亚型。     

pH值对猪场养殖废水常温短程硝化特性的影响

为实现高氨氮猪场养殖废水高效脱氮处理,针对猪场废水中尿素(主要源于猪尿液)水解过程伴随着污水pH值的变化现象,采用序列间歇式活性污泥法(SBR)处理某猪场养殖废水,在常温(20℃)、非限制溶解氧的条件下,考察了pH值、曝气时间等因素对氨氮去除效果和短程硝化特性的影响.实验结果表明,pH值在7.0 ～9.5范围内亚硝酸盐积累率在75％～80％之间;长时间曝气并未对短程硝化造成影响;高质量浓度氨氮废水生物处理过程中,亚硝酸盐积累与进水pH值、游离氨(FA)及游离亚硝酸(FNA)有关.     

序批式反应器处置打捞蓝藻的初步研究

[目的]研究序批式反应器对蓝藻的降解规律。[方法]采用序批式反应器好氧-厌氧资源化处置蓝藻,分析蓝藻中蛋白质、水溶性多糖、藻毒素的含量变化及甲烷产量。[结果]蓝藻中蛋白质和水溶性多糖类降解率较高,而且在好氧阶段完成大部分降解。藻毒素降解率为45%;沼气在厌氧60 d时,沼气体积占收集容器体积的62.1%。[结论]该研究为蓝藻的综合处置及利用提供了理论依据。     

Breeding and biotechnological efforts in Jatropha curcas L. for sustainable yields

Jatropha is a non-edible, important bioenergy plant, which can grow in marginalized land. The seeds possess about 36% oil and this would be converted into biodiesel or biojet-fuel. Jatropha provides an option for sustainable feed and fuel production due to its inherent qualities including hardy nature, drought tolerance and surviving with limited amount of water, tolerance to unfavorable conditions and excessive moisture. However, heterozygosity, low productivity and poor understanding of its genome are the major impediments to elite line development. Further, classical breeding and advanced technological investments remain limited owing to long juvenile phase and breeding cycles. Scientific technologies that lead to identification of elite genotypes and development of high yielding elite Jatropha lines and effective methods of detoxification of seeds needs immediate priority. Efficient tissue culture system, doubled haploids (DH) and genomic tools are increasingly made available to improve the seed yield and its oil quantity through the development of geminivirus disease resistant lines. The application of advanced, sequencing technologies has presented a repertoire of genomic information for this important yet orphan crop. In the present investigation, we highlight the achievements made in Jatropha towards development of high yielding, virus resistant elite lines and hybrids with yield potential ranging from 3 to 5 tons per hectare in a year, which is a first ever report in the world. We also developed potential biotechnological tools such as genetic transformation, genome editing and next generation genomics tools including linkage maps and QTLs for accelerating breeding efforts through marker assisted selection. Because of our concerted and continuous efforts during the past 15 years, we have overcome all the obstacles and developed high yielding, disease resistant hybrids/lines, advanced cultivation technologies with thorough knowledge on Agronomy of the Jatropha crop. The Jatropha cultivation technology developed for the first time in the world, could open up new avenues for higher yield productivity of commercial viability.     

猪繁殖与呼吸综合征病毒BJ株、GD株全基因组序列测定与分析

利用RT-PCR方法分别扩增出猪繁殖与呼吸综合征病毒(PRRSV)北京分离株(BJ株)、广东分离株(GD株)各9条基因片段,并将这9个片段分别克隆于PGEM-T-easy质粒载体上进行测序,按顺序将这些序列进行拼接得到PRRSVBJ株和GD株的全基因组cDNA(GenBank:EU825723,EU825724)。测序结果表明,BJ株和GD株基因组全长分别为15340和15336bp,各包含9个开放式阅读框,5′UTR都含有189nt,3′端URT分别含有170nt、166nt,其中各包含20ntPoly(A)、16ntPoly(A)。基因组序列分析结果显示,BJ株和GD株与ATCCVR-2332的核苷酸同源性分别为88.9%和89.0%,与高致病性PRRSV毒株JXA1的核苷酸同源性分别为99.0%和99.4%,与PRRSV毒株HuN的核苷酸同源性分别为98.8%和98.9%。     

毛竹β-1,3-葡聚糖酶基因的克隆及序列分析(英文)

β-1,3-葡聚糖酶是一种植物病程相关蛋白,在植物抵御病害中有重要作用。本研究以毛竹为实验材料,利用RACE技术,克隆毛竹β-1,3-葡聚糖酶基因的外显子序列,并在β-1,3-葡聚糖酶基因的编码区两端设计引物,以毛竹基因组DNA为模版扩增该基因的内含子序列。序列结果表明,该基因全长1693bp,包含两个外显子和一个内含子,命名为PheGLU(GenBank登录号GU238236)。该基因包含1个996bp的开放阅读框,编码332个氨基酸,相对分子质量为3.541×104Da,其等电点pI为6.389,是一个酸性蛋白且分泌到胞外;其中,该蛋白的二级结构中包含35.15%的α-螺旋,21.82%的β-转角,43.03%的延伸链和无规则卷曲等;三级结构同源建模预测显示,它与大麦β-1,3-葡聚糖酶(PDB number:1ghsA)具有80.4%的同源性;聚类分析显示,该基因序列与已报道的其它植物具有较的高氨基酸序列同源性。本研究为进一步鉴定毛竹β-1,3-葡聚糖酶基因的抗真菌病害能力奠定了基础。     

猪核转录因子C/EBPβ基因3′末端部分序列的克隆

[目的]研究猪核转录因子C/EBPβ,扩增C/EBPβ3′非翻译区（3′UTR）部分序列。[方法]以报道的猪C/EBPβ基因片段（DQ450678.1）为模板设计引物,应用3′末端快速扩增技术（3′RACE技术）进行扩增。[结果]成功克隆猪核转录因子C/EBPβ的3′UTR,并获得GenBank登录号：FJ869121。[结论]与GenBank中报道的其他哺乳动物相应序列进行比较分析,猪核转录因子C/EBPβ的3′UTR序列与人C/EBPβ基因核苷酸序列BLAST比对,同源性为93%,为克隆其基因组全长及分析其功能奠定了基础。     

丰田混合型装配线的排序方法

给出了丰田装配线平稳生产的2个目标,提出了混合型装配线上的作业时间与循环周期的表达式,指出了丰田排序算法的基本概念在于缩小生产过程的零件平均消耗速度与实际消耗速度的差异,总结了丰田恒定零件消耗速度目标追逐法的步骤。根据某产品的产量及其所需零件数目推导了它的生产排序进程,指出了当零件数目较多时,以时间段为单位来调整零件消耗速度。     

Genetic variability of <Emphasis Type="Italic">Plum pox virus</Emphasis> isolates in the Czech Republic

Plum pox virus (PPV), the causal agent of Sharka disease, is an important pathogen of stone fruit trees. In this study, 24 new Czech PPV isolates from five different orchards were collected and characterized, molecularly. PPV-D isolates were identified in all orchards studied; whereas PPV-Rec isolates were identified in only two of them. A phylogenetic analysis on (Cter) NIb-(Nter) CP was performed. Three Czech PPV-D isolates BOH11CZ, BOH12CZ, and BOH13CZ diverged into a significantly separated cluster. PPV-Rec isolates formed a fairly homogenous group. However, the Bohutice and the Lipov PPV-Rec isolates clustered in two significantly separated branches.