克隆

The Omp25 gene of wapiti's Brucella abortus was amplified by RT-PCR and sequenced, Then the interesting gene was subcloned into pET28a for expression. The results showed that the full length of Omp25 gene was 642bp, which encoded 213 amino acids.

运用RT-PCR技术从马鹿布氏杆菌分离株中扩增出Omp25基因，将其克隆入pMD18-T中进行核苷酸序列测定和遗传变异分析，并将目的基因亚克隆到大肠杆菌表达载体pET28a中诱导表达。

The results showed that there were significant differences in numbers of stolons, length of stolons, dry weight of genets, dry weight of stolons and dry weight of ramets of L. brachystachya in different plots with different altitudes (P.01). The numbers of stolons of L.

在一定时间内，克隆植物相互连接的克隆分株可生长在不同环境条件的斑块中，并彼此进行物质交换、实现克隆整合，从而最大可能地利用异质环境 [5]。

Vulgaris and Beta M14 BAC clone 16-M11 and 26-L15 are florescence specific expression BAC of Beta M14, both BAC clone 16-M11 and 26-L15 were in situ hybridization to monosomic addition lines(2n=2x=18+1) M14 chromosomes, they were located on the long arm of corollinae chromosome 9. the dominant factor of BAC clone 16-M11 and 26-L15 appears to hemizygous, because no equivalent region has been found in sexual Beta.

选用这两个BAC克隆作为探针，对M14甜菜有丝分裂中期染色体组进行荧光原位杂交，结果发现，BAC克隆16-M11和26-L15定位于M14甜菜附加的一条白花甜菜9号染色体上，有性栽培甜菜染色体上没有出现信号，两个BAC克隆在M14甜菜中呈半合子状态，遗传行为象单价显性基因，是白花甜菜9号染色体特异的。

Using the total RNA of merozoites of Eimeria acervulina isolated from Guangdong Pro- vince as template,a partial segment of 3-1E gene was amplified by RT-PCR. The gene fragment 682 bp inlength was cloned into pGEM-T Easy vector,and the recombinant plasmid was identified by PCR,restric-tion enzyme analysis and sequencing. The homology analysis revealed that the nucleotide homology be-tween the 3-1E gene of the E.

根据GenBank中登录的堆型艾美球虫3-1E基因序列，设计了3条引物，以广东株堆型艾美球虫裂殖子总RNA为模板，利用反转录-聚合酶链反应扩增获得了3-1E基因部分片段，将这一片段克隆至pGEM-T Easy载体中，经PCR、限制性内切酶分析和克隆片段的序列测定、比较，证实了克隆片段的可靠性。

The results show that DCRP is indeed a excellent flame retardant. The material can reach the UL-94-V-0 level through choice different synergist. Many alternative synergists to antimony oxide are available for use. Zine and iron compounds are often used in place of the standard antimony oxide because of their lower cost and special properties. Additionally, synergist mixtures are also useful in obtaining specific properties.

本文在参考国内外文献的基础上自行设计了一些配方，分别把得克隆应用于尼龙66、尼龙6、PBT、PP、ABS等材料中，对得克隆进行了应用研究实验，发现得克隆可赋予材料优异的阻燃性，通过选用不同的协效剂一般可以达到UL-94-V-0阻燃级别，并且，可供选择的协效剂不仅是三氧化二锑，还有多种其他化合物如氧化铁、硼酸锌等及它们的配合物，这类协效剂在某些方面还优于三氧化二锑。

The results show that DCRP is indeed a excellent flame retardant. The material can reach the UL-94-V-0 level through choice different synergist. Many alternative synergists to antimony oxide are available for use. Zine and iron compounds are often used in place of the standard antimony oxide because of their lower cost and special properties. Additionally, synergist mixtures are also useful in obtaining specific properties.

本文在参考国内外文献的基础上自行设计了一些配方，分别把得克隆应用于尼龙66、尼龙6、PBT、PP、ABS等材料中，对得克隆进行了应用研究实验，发现得克隆可赋予材料优异的阻燃性，通过选用不同的协效剂，一般可以达到UL-94-V-O阻燃级别，并且，可供选择的协效剂不仅有三氧化二锑，而且还有多种其他化合物如氧化铁、硼酸锌等及它们的配合物，这类协效剂在某些方面还优于三氧化二锑。

The cell cultures were monitored by provirus DNA PCR, RT-PCR, reverse transcriptase activity assay and real-time RT-PCR, The results of RT activity and DNA PCR, RT-PCR were positive in the supernatant of cell cultures and viral particles were also clearly observed under electron microscope.

本试验基于代次毒分析结果，选取LTR 变异率较高的U3 区，以驴胎皮肤弱毒疫苗感染性分子克隆株pLGFD3-8 和驴强毒株感染性分子克隆株为父本操作系统进行基因替换，构建EIAV 非编码区强弱毒嵌合的全基因分子克隆，将阳性重组质粒命名为pLGFD9-12。

Nested PCR analysis and southern hybridization analysis showed that no polymorphism between H.villosa, Pm97034 and susceptible parent Wan7 107 was detected with the clones from 6VS arm, whereas three clones from H.villosa genome DNA: RH42, RH55, and RH66 showed polymorphism. RGA6 cloned from H.villosa genome DNA was characterized identity to NBS, and show high homology to resistance genes as RPM1 and RPP13 in Arabiadopsis, LRR19 in wheat, and I2C-1 in tomato.cDNA library constructed from T.aestivum-H.villosa translocation line Pm97034 was screened by hybridization with RGA6. Four positive cDNA clones were obtained. R3-2-2 showing 60-90% identity with eukaryotic sulfite oxidases contained an open reading frame of 647bp encoding 140 amino acid, and contained a conserved Moco-dimer domain in the ORF. R6-2-2 showed an ORF containing an Euk-porin domain of 279aa with 20-40% identity to the eukaryotic voltage-dependent anion channel proteins. R8-1-1 showed a complete ORF of 1810bp encoding 493aa with 80-90% identity to plant catalases. R9-1-1 showed an ORF containing a BTB/POZ conserved domain of 204aa with 30-70% identity to pox virus and zinc finger proteins. R3-2-2 and R9-1-1 were the first cDNA clones containingconserved domain of SO and POZ respectively isolated from wheat. R8-1-1 contained a complete ORF with 81% identity to Cat-3. Aspects of the role of R8-1-1 may be same with Cat-3, and it would offer the opportunity for improvement of stress tolerance of wheat.

以RGA6为探针，筛选用抗白粉病小麦—簇毛麦易位系Pm97034构建的cDNA文库，得到了4个阳性克隆。R3-2-2与动植物的亚硫酸氧化酶(sulfite oxidase，SO)有60～90％的同源性，长度为647bp，编码140个氨基酸，具有开放阅读框并含有保守域Moco-dimer.R6-2-2与真核生物的VDAC(voltage-dependent anion chennel)蛋白有20～40％的同源性，长度为1047bp，编码279aa，是一个不完整的开放阅读框，预测结构具有Euk-porin结构域。R8-1-1与植物中已经克隆的过氧化氢酶有80～90％的同源性，长度为1810bp，编码493aa，具有完整的开放阅读框和过氧化氢氧化酶保守域。R9-1-1与动植物的POZ(pox virus and zinc finger protein)蛋白有30～70％的同源性，长度为1446bp，具有开放阅读框和BTB／POZ保守域。R3-2-2与R9-1-1是首次从小麦中克隆到的具有SO和POZ保守域的cDNA序列。R8-1-1具有完整的开放阅读框，与玉米Cat-3基因具有81％同源性，预测R8-1-1可能具有与Cat-3类似的功能，可为转基因小麦抗逆育种提供新的基因。

New laccase gene and its full-length cDNA were clone from genomic library and equalized cDNA library of Ganoderma lucidum. Similarity between the sequence and other laccase gene were analyszed with NCBI blast, the results show that the new gene belong to multicopper oxidases gene family and have 60-70% Similarity with other laccase gene.

从灵芝的基因组文库和cDNA均一化文库中克隆了灵芝的漆酶基因和全长cDNA，并通过NCBI Conserved Domain Search氨基酸序列的相似性分析及与其它真菌漆酶之间的相似性分析，证明得到的蛋白是一种新的multicopper oxidases，其蛋白质序列与其它已克隆漆酶基因的同源性在6 0～7 0％之间，并对论文克隆的漆酶基因的启动子和终止子进行了分析。

This research is taking sea perch as the material, and cloning the Syntaxin 1B gene. After extracting RNA from sea perch tissue, we gained the first chain of cDNA by RT-PCR. Then designing a pair of primers according to the sequence of Syntaxin 1B from other organism. Based on the first chain of cDNA cloned from sea perch, the partial sequence of Syntaxin 1B gene is been cloned by using TD-PCR and we received the 751bp fragment at last.

本实验从鲈鱼脑组织中提取总RNA，通过反转录得到cDNA第一条链，根据已克隆出的其它生物的Syntaxin 1B基因序列，设计并合成引物，在妒鱼syntaxin1B基因部分序列克隆与分析 cDNA第一条链的基础上使用降落PCR，进行体外扩增妒鱼 Syntaxin IB基因的部分序列，得到长度为75 lbP的片断，在胶中洗脱下该片段，与pMD 18一T载体链接，转化入感受态细胞，挑取阳性克隆扩大培养，提取质粒，经HindIH和x一gaH两内切酶消化得到75 1 bp的片断，证明链接成功。

With Death guitarist Schuldiner adopting vocal duties, the band made a major impact on the scene.

随着死亡的吉他手Schuldiner接受主唱的职务，乐队在现实中树立了重要的影响。

But he could still end up breakfasting on Swiss-government issue muesli because all six are accused of nicking around 45 million pounds they should have paid to FIFA.

不过他最后仍有可能沦为瑞士政府&议事餐桌&上的一道早餐，因为这所有六个人都被指控把本应支付给国际足联的大约4500万英镑骗了个精光。