基因文库

Results (1) The forward and reverse subtracted cDNA libraries of different metastastic potential large cell lung cancer cell lines were successfully constructed;(2) With blue-white colony and dot blot, 307 positive clones in the forward subtracted library and 78 positive clones in the reverse subtracted library were obtained;(3) 55 clones were successfully sequenced in the forward subtracted library. Homolog analysis confirmed 23 differential expression segments, which were similar to the human genes already known, including NME2, NPM1, MT 2A, HSPE1, TAFIA, EPRS, PX19 and EIF3S9 et al;(4) 31 clones were successfully sequenced in the reverse subtracted library. Homolog analysis confirmed 16 differentially expressed segments. 15 of them were similar to the human genes already known, including ANXA2, TUBB, PKN2, GNAS, EEF1A1, SSR2 and RPLPO et al. Only one segment had partial homology to known human genes. This segment was supposed to be the new EST segments which might not have been cloned.

结果(1)成功构建人大细胞肺癌高低转移株差异表达基因正向消减cDNA文库和反向消减cDNA文库；(2)经蓝白菌落筛选和斑点杂交，正向消减文库获得307个阳性克隆，反向消减文库获得78个阳性克隆；(3)正向消减文库挑选55个克隆成功测序，经同源性分析最终确定差异表达基因片段23个与已知人类全长基因具有很高相似性(95％～100％)，这些基因包括NME2、NPM1、MT2A、HSPE1、TAFlA、EPRS、PX19和EIF3S9等；(4)反向消减文库挑选31个克隆成功测序，经同源性检索和比对最终确定差异表达基因片段16个，其中15个与人类全长基因具有很高相似性(95％～100％)，包括ANXA2、TUBB、PKN2、GNAS、EEF1A1、SSR2和RPLPO等基因。1个片段和己知人类基因仅有部分相似性，表明它可能是未被克隆的人类基因EST片段。

Using semi-quantitative RT-PCR, the differential expression profiles of eleven selected genes were confirmed in the ovaries of triploid and diploid. These genes fell in gene categories with a wide range of functions. The results indicated that triploidy affects the dynamic gene regulatory network in triploid ovary. This study established a firm basis for future investigation on characterization of crucial molecular events for normal ovarian development in shrimp.To further dissect exact gene functions for gonad development of shrimp, three differentially expressed genes between diploid and triploid ovary, PCNA (proliferating cell nuclear antigen), CAS/CSE1 (cellular apoptosis susceptibility protein/chromosome segregation 1) and SSRF (spermatogonial stem-cell renewal factor) were characterized on certain aspects.

利用抑制性消减杂交技术，建立了对虾二倍体和三倍体卵巢间的2个消减文库；在正向消减文库(以三倍体卵巢作为实验组，二倍体卵巢作为驱动组)中，鉴定到54个基因；在反向消减文库(以二倍体卵巢为实验组，三倍体卵巢为驱动组)中，鉴定到16个基因；选取11个差异表达的基因，利用半定量RT-PCR的方法对其在二倍体和三倍体卵巢间的表达进行了检测，均能很好地与消减结果相吻合；这些差异基因编码多种功能的蛋白，分析表明染色体的三倍化使三倍体卵巢中的基因调控网络受到了影响；为深入揭示维持卵巢正常发育的关键分子调控事件奠定了基础。

Then we transformed mTn-lacZ/LEU2 transposon library into the YES2MTM1 strain. Transformants lost the plasmid overexpressing MTM1 after 5-Fluoroorotic acid treatment. We picked up the yeast strains able to grow on nonfermentable carbon source with MTM1 deletion and some transposon insertion and identified the insertion sites.

因MTM1基因的缺失造成的损伤不可逆，直接转入文库无法筛选得到MTM1 基因缺失表型相关基因，本研究利用外源MTM1基因菌株和mTn-lacZ/LEU2酿酒酵母转座子文库进行筛选，寻找能挽救mtm1突变体生长缺陷的转座子插入位点。

Results A total of 121 positive clones were randomly screened from the library and 22 Operational Taxonomic Units were determined. BLAST analysis indicated that all OTUs were affiliated with the phylum Crenarchaeota.

从冷泉沉积物古菌16S rRNA 基因文库中随机挑选了121个阳性克隆，共得到22个不同的可操作分类单元，BLAST结果表明全部克隆子归属于泉古菌门中免培养类群。

Functionally known genes were obtained after Blastx analysis, 237 of them were from the subtractive library of stage of proembryo mass and 279 from that of somatic embryo maturation.

成功构建了Y35体细胞胚原胚团阶段和成熟阶段的差异表达基因文库，获得了868个差异表达的UniESTs，其中原胚团阶段400个，体细胞胚成熟阶段468个。

A strain NK13 was screened for a certain extent asymmetric hydrolyze the rac-ketoprofen Chloroethyl ester and identified as Bacillus megaterium. For the preparation of gene libraries, a positive clones was obtained from the tributyrin flat. The sequence of this esterase gene had been analysised, and contained the whole ORF of an esterase gene with the length of 933bp.

以本实验室筛选出的一株具有不对称拆分消旋酮基布洛芬氯乙酯的菌株NK13为材料，经初步鉴定为巨大芽孢杆菌，通过构建其基因文库，从中筛选得到一阳性克隆重组子pUC-NK。

Speculating on the figure of two dimensional electrophoresis of rice glutelins there are at least 13 glutelin genes, only six of which, GluA-1, GluA-2, GluA-3, GluB-1, GluB-2 and a cDNA clone,λ RG21 cDNA, have been sequenced.

从水稻谷蛋白的2-D电泳可推测，编码水稻谷蛋白的基因至少有13个，目前尚只有6个水稻谷蛋白基因被分离测序，它们分别是GluA-1，GluA-2，GluA-3，GluB-1，GluB-2和一cDNA克隆（λRG21 cDNA），而且，这些基因都是通过实验筛选大量的水稻基因组文库或cDNA文库获得的，其过程成本高，工作量大，实验周期长。

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。

Dendritic cells are the most potent antigen-presenting cells in human immune system. Our institute has been working on the region of DC development and DC-mediated immune therapy on cancers for a long time. A cDNA library of human dendritic cells has been set up and by large-scared random sequencing, we have isolated a lot of novel fulllength cDNA clones.

树突状细胞(Dendritic cells，DC)是人体内重要的抗原提呈细胞，本研究所对DC的分化发育及其介导的肿瘤免疫治疗的研究工作已进行多年，并在此基础上建立了人树突状细胞基因文库，通过大规模随机测序的方法开展了从DC中克隆新分子的工作，从2万余条EST中发现了许多新的分子。

Clarke-Carbon equation can give a good estimation for the necessary number of recombinants in a gene library.

Clarke-Carbon公式用于估计一个完整的基因文库所需的最少克隆数。

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。