克隆

Tuberculosis H37Rv strain was amplified by PCR and cloned to vector pEASY T1. After sequencing, it was subcloned to fusion expression vector pET28a and then transfected to E coli BL21 (DE3). It was found that the LprG protein was successfully expressed after induction with LPTG.

采用聚合酶链反应从结核分枝杆菌H37Rv基因组中扩增出LprG基因，克隆到pEASY T1中，序列测定正确后，再亚克隆到融合表达载体PET28a中，转化入大肠杆菌BL21(DE3)，经IPTG诱导表达。

Currently, neither cloning S and Z gene nor identifying their products was successful. Phalaris coerulescens can be used as a model plant to study SI in the Poaceae. Segregating population and comparative genomics strategy were used to map S and Z, constructing high-resolution mapping of the target loci so that map base clone S and Z genes. It is a research orientation and hot spot of self-incompatibility in Poaceae.

目前S、Z基因的克隆和编码产物的鉴定均未有成功的报道，天蓝虉草可作为禾本自交不亲和性研究的一个模式作物，运用分离群体和比较基因组学方法构建S与Z位点的精细连锁图谱，以对自交不亲和基因进行图位克隆，是当前禾本科植物自交不亲和性研究的一个方向和热点。

For the substitution and transversion of nucleotide in the same family, CHS sequences from Poaceae, Solanaceae and Papilionoideae evolved at nearly the same rate.

我们从水稻的基因组BAC文库中克隆了2个CHS基因，即B1-P7和B2-P5.B1-P7包含有完整的CHS基因，B2-P5则只克隆了外显子2和3'端的非编码区。

In attempt to elucidate the molecular mechanism of cold tolerance of wintersweet, which blooms in the deep winter,a COR413 protein gene, designated as Cpcor413pm1, was obtained by sequencing the randomly selected clones, on the basis of Chimonanthus praecox flower cDNA library construction and its ESTs analysis.

为研究蜡梅冬季开花过程的抗寒分子机理，在构建蜡梅花cDNA文库及EST分析的基础上，通过随机克隆测序，克隆了1个蜡梅COR413蛋白的cDNA基因，命名为Cpcor413pm1。

RESULTS: On the basis of either flow-cytometric or molecular analysis, 44 of 45 patients with CLL (98%; 95% confidence interval , 88 to 100) had a prediagnostic B-cell clone; in 41 patients (91%; 95% CI, 79 to 98), the presence of the B-cell clone was confirmed by both methods. The presence of immunoglobulin heavy-chain variable genes was determined in 35 of 45 prediagnostic clones (78%).

结果：在流式细胞或分子分析的基础上， 45名白血病患者有44例（98 ％； 95 ％可信区间，88～100 ）具有诊断前B细胞克隆；在41例（ 91％； 95 ％可信区间， 79～98 ），B细胞克隆的存在得到了这两种方法的证实。45个诊断前克隆中有35个(78%)具有免疫球蛋白重链可变基因。

This may be mostly resulted from the smaller lateral root diameter and the nearer distance between clonal ramet and mother plant or related to the environmental conditions of different population patches or different management measures.

可能是产生克隆分株的根径较小和克隆分株距离母体植株较远造成的，同时也可能与不同种群斑块环境条件的差异或不同的管理措施有关。

Methods: MUC1/Y extracellular domain was used as a target molecule to biopan Ph. D. 12 phage randem peptide library. Two protocols using affinity gel and cell culture plates respectively were carried out. Positive phage clones were identified by ELISA. ssDNA sequencing was done on 16 positive phage clones to get the amino acid sequences of MUC1/Y-binding peptides. Immunohistochemistry was done to show the capacity and specificity of positive phage clones to bind the tumor cell lines.

以MUC1/Y黏蛋白的胞外段蛋白(MUC1/Yex)为靶分子，用凝胶亲和法和酶联板法分别筛选十二肽噬菌体随机肽库，ELISA鉴定阳性克隆，DNA序列测定后确定MUC1/Yex结合肽的氨基酸序列；免疫组化鉴定阳性噬菌体克隆与正常及肿瘤细胞的结合能力及特异性。

Methods: Total RNA was extracted from esophageal carcinoma tissue, the primer containing special enzyme was designed, NY-ESO-1 fragment was amplified by RT-PCR, and then was cloned into the expression vector pET-15b by LP Recco PCR Cloning. The recombinants plasmid pET-15b-NY-ESO-1 were identified by restriction endonucleases digest analysis and sequence analysis.

从人新鲜的食管癌组织中提取总RNA，通过RT-PCR技术扩增NY-ES0-1基因3'端的340bp片段，采用靶向克隆法将目的基因插入到原核表达载体pET-15b中，得到重组表达质粒pET-15b-NY-ESO-1，经过筛选，挑选出阳性克隆进行XhoⅠ和BamHⅠ双酶切图谱分析、PCR检测和扩增产物核苷酸序列分析等，鉴定所构建的原核表达载体。

MethodUsing the canis MC4R DNA as template,the specific primers were designed. After PCR amplification,product was cloned into pGEX-4T-1 vector by LP Recco PCR cloning technique,the recombinant pGEX-4T-1-cMC4R was transferred to E.

方法以Beagle犬基因组DNA为模板，经PCR技术扩增目的片段，利用LP Recco PCR克隆技术将目的片段直接重组到原核表达载体pGEX-4T-1上，转化到E.coli DH5α，筛选阳性克隆，BamHI、XhoI酶切鉴定，DNA 测序检测插入序列的正确性。

During the process of nuclear reprogramming, the zygotes of the normal embryos and the donor nuclei of thecloned embryos cease their unique repertoire of gene expression and reset their gene expression to the totipo-tent status, and then the normal and cloned embryos redifferentiate from the totipotent status to variousdifferentiated states for tissue generation or organogenesis.

通过细胞核重编程，首先，受精卵和克隆胚胎的供体核停止其特有的基因表达程序，恢复为全能状态的基因表达程序；然后，受精胚胎和克隆胚胎的细胞再从全能状态重新进入分化状态，最终形成各种组织和器官。

Lugalbanda was a god and shepherd king of Uruk where he was worshipped for over a thousand years.

Lugalbanda 是神和被崇拜了一千年多 Uruk古埃及喜克索王朝国王。

I am coming just now,' and went on perfuming himself with Hunut, then he came and sat.

我来只是现在，'歼灭战perfuming自己与胡努特，那麼，他来到和SAT 。