克隆

So it couldn't fit dynamic network environments soundly. After investigating of negative selection, affinity mature and memory detector evolution mechanisms, it is Embedded Negative Selection Operator Clonal Selection Algorithm presented combined with negative selection, clonal selection and memory detector gene library evolution.

本文结合网络入侵检测的上述问题，深入研究了否定选择、亲和力成熟过程以及免疫记忆机制等，以现有克隆选择算法为主体，将否定选择、克隆选择、记忆检测器基因库方法融合进来，提出嵌入否定选择的克隆选择算法(Embeded Negative Selection Operator Clonal Selection Algorithm， ENCSA)。

A clone from each genotype was randomly selected as representative for sequencing. The obtained 16S rDNA gene sequences had a similarity of 87%-100% with those in the GenBank (www. ncbi. nlm. nih. gov), and more than half of them had a similarity lower than 97%, being of new species. Based on phylogenetic analysis, the bacteria in the two soils were classified into 10 groups, with 5 groups in common. The dominant bacterial groups in the two soils differed obviously. In primeval forest soil, the dominant group was Proteobacteria, which had 39 genotypes, occupying 58.0% of all the clones; while in the soil of degraded ecosystem the dominant groups were Acidobacteria and Proteobacteria, which had 19 and 15 genotypes occupying 32.5% and 30.5% of all the clones, respectively. In the soil of degraded ecosystem, Proteobacteria group decreased while Acidobacteria group increased markedly, compared with those in primeval forest soil.

从每种基因型中随机选择一个克隆子作为代表进行测序分析，所有序列与GenBank数据库中序列的同源性为87%~100%，且两样地中均有超过一半的基因型序列与数据库中已知序列同源性低于97%，属于分类在"种"地位上的新发现细菌；通过系统发育研究将两样地的细菌分为10大类群，两样地共同拥有5大类群，但两样地的细菌优势类群明显不同，原生土壤为Proteobacteria，含39种基因型，占总克隆子数的58.0%，退化生态系统土壤为Acidobacteria和Proteobacteria，分别含19种和15种基因型，占总克隆子数的32.5%和30.5%；与原生土壤细菌类群相比，退化生态系统土壤Proteobacteria类群明显减少，Acidobacteria类群明显增加。

Producing PHAs,and then were orientationally inserted into a prokaryotic expression vector pBV220. By analyzing the bioactibity and function of their products, the biological function of phaA, phaB and phaC was confirmed. These have been done in our laboratory. To make better use of the three genes by plant bioreactor, two groups of different plant expression vectors that can be used to produce PHAs in different plants were constructed, and the transformation of PHA in Oxalis triangnlaris has been preliminarily reseached.The following is the main results:Three plant expression vectors pBI121-A,pBI121-B and ubquiting-C, which can be used to be transformed into Oxalis triangularis were respectively constructed by using the phaA, phaB and phaC.

菌株的亚克隆基因组片段中，分离出phaA、phaB和phaC三个基因片段，定向克隆至原核表达载体pBV220，通过对其生物活性与功能分析，确证了基因phaA、phaB和phaC的生物学功能；为了通过植物生物反应器对这三个基因进一步研究利用，本实验利用这三个基因构建了两组可用于不同植物生产PHA的真核表达载体，并对紫叶酢浆草进行PHA基因遗传转化的初步研究，获得主要结果如下：利用克隆的phaA、phaB、phaC三个基因分别构建了可用于转化紫叶酢浆草表达载体pBI121-A、pBI121-B、uBquiting-C；可用于玉米转化的种子特异性表达载体p7S-A、pBI121-S-B、uBquiting-C。

The PCR product was cloned into pCR BluntⅡ-TOPO vector to form subclone and then sequenced.The subclone product were cloned into the improved pBluescript Ⅱ KS vector in proper order by suitable enzyme sites to obtain viral full length genomic cDNA clone.

根据测序结果，选择合适的酶切位点将各个亚克隆产物逐段克隆入改造过的pBluescript Ⅱ KS载体中，从而获得了病毒的全长基因组cDNA克隆。

There were some differences in responses of life history characteristics to increasing temperature among the four rotifer clones.

四温度间，克隆A轮虫幼体的耐饥饿时间在15℃下最长，克隆B和C轮虫幼体的耐饥饿时间在30℃下均最短，克隆D轮虫幼体的耐饥饿时间随培养温度的升高而逐步显著地缩短。

PCRs were performed on genomic DNAs of four pear cultivars Sand pear, Huangguan pear, Xueli pear, Yali pear using primers derived from sequences of previous published PPO. The target bands were recovered and purified. The fragments were ligated with the plasmid of pGEM-T Vector by the method of direct T-A cloning.

以4种不同来源的梨基因组DNA为模板，根据已经发表的多酚氧化酶基因序列设计引物，利用PCR技术，克隆到了鸭梨、雪花梨、砂梨、黄冠梨约1.8 kb的完整多酚氧化酶基因，将纯化后的扩增产物克隆到质粒pGEM-T vec-tor中，转化DH-5α菌，挑选阳性克隆进行测序。

The total RNA was abstracted from the larvae of Boophilus microplus, and a 1982bp Bm86 gene was amplified by RT-PCR. The target gene was subcloned into T vector. The sequencing showed that the nucleotide sequence of the cloned Bm86 gene shared 97.4% homology with the data published in and this fragment contained the complete open reading frame of Bm86 gene.

为了克隆微小牛蜱Bm86 基因及构建该基因的表达载体，以微小牛蜱饥饿幼蜱的破解物提取的总RNA为模板，参照已发表的微小牛蜱Bm86基因的核苷酸序列，设计了1对引物，采用RT-PCR技术获得微小牛蜱Bm86基因；将Bm86基因克隆入载体，并进行序列分析，结果证明，克隆的Bm86基因序列与GenBank上登录的Bm86基因序列的同源性达97.4%，而且该序列包含完整的开放阅读框。

ASES is 59% identical to Artemisia cyclase cDNA clone cASC125, 50% identical to epi-cedrol synthase from A. annua, 48% identical to amorpha-4, 11-diene synthase from A. Annua, 39% identical to the 5-epi-aristolechene synthase from tabacco, 38% identical to vetispiradiene synthase from H. Muticus, 41% identical to the δ-cadinene synthase from cotton. The coding region of cDNA was cloned into procaryotic expression vector pET30a and overexpressed in E. coli BL21 (DE3). The cyclase proteins extracted from bacterial culture were found largely in the insoluble protein fraction. The tissue specific expression patterns were analyzed by RT/PCR.

克隆的倍半萜合酶氨基酸序列与烟草马兜铃烯合酶、莨菪岩兰螺旋二烯合酶、棉花杜松烯合酶的一致性分别为39％，38％和41％；与青蒿柏木脑合酶、紫穗槐二烯合酶和一个推测的倍半萜合酶克隆cASC125的一致性为50％，48％和59％。cDNA编码区序列被克隆进原核表达载体pET-30a，并在大肠杆菌BL21（DE3）中诱导表达，但过量表达的蛋白主要是以不溶性蛋白形式存在。

Methods Totally 173 patients diagnosed as MDS according to FAB or WHO criteria with complete clinical and cytogenetical data were included in this reseach.

方法应用骨髓细胞短期培养法和染色体G显带技术，部分病例联合应用荧光原位杂交技术，对42例细胞遗传学检查具有+8异常克隆，16例具有-7/7q-异常克隆，以及55例虽然经常规细胞遗传学检查未检出异常克隆，但是骨髓涂片原始细胞≥0.10，按照FAB或WHO标准既往诊断为MDS的病例进行了临床及血液形态学和细胞遗传学的系列研究。

Methods Totally 173 patients diagnosed as MDS according to FAB or WHO criteria with complete clinical and cytogenetical data were included in this reseach. Among them 42 had +8 chromosome aberration, 16 had -7/7q-, and 55 had normal karyotypes and more than 0.10 blast cells in the bone marrow. Short term culture and G-banding techniques and in some specimens fluorescence in situ hybridization method were used to do chromosome analysis.

方法应用骨髓细胞短期培养法和染色体G显带技术，部分病例联合应用荧光原位杂交技术，对42例细胞遗传学检查具有+8异常克隆，16例具有-7/7q-异常克隆，以及55例虽然经常规细胞遗传学检查未检出异常克隆，但是骨髓涂片原始细胞≥0.10，按照FAB或WHO标准既往诊断为MDS的病例进行了临床及血液形态学和细胞遗传学的系列研究。

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。