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Results The eukaryon expression system pcDNA3.1/myc-HisC-bFGF could express the target protein bFGF in vitro. The recombinant plasmids, pcDNA3.1/myc-HisC-bFGF and pCD2-VEGF121 were transferred into muscles flap in vivo successfully. The active proteins bFGF and VEGF121were expressed at high levels.

结果 构建的pcDNA3.1/myc-HisC-bFGF真核表达体系成功转染体外培养HeLa细胞，目的基因在mRNA水平和蛋白水平均有表达。pcDNA3.1/myc-HisC-bFGF和pCD2-VEGF121重组质粒分别转染在体肌瓣，获得外源基因高水平表达。

Pestis pathogenicity. Methods: We utilized "whole genome shotgun" approach to get the genome sequence of 91001. Based on the finished and annotated genome sequence of 91001, as well as previously published genome sequence of C092 and KIM, we performed detailed comparative genomics analysis on their chromosome and plasmids. For the important phenotypes - nitrate reduction and glycerol fermentation, we carried out large scale screening among 257 strains isolated in China to clarify the genetic mechanism of these two phenotypes.

采用全基因组鸟枪法测定91001菌株的全基因组序列，利用比较基因组方法对91001与另两株鼠疫耶尔森氏菌(CO92和KIM)的全基因组进行比较研究；同时针对一些有意义的基因在多株鼠疫耶尔森氏菌的中国分离株中进行了验证；并在基因组研究的基础上开展了初步的蛋白质学研究。

Cell lines were co-tranfected with bi-plasmids and cultured at 1% O〓 condition, luciferase assay demonstrated that all of the expression systems with different domains could be hypoxia-activated. GAL-CAD achieved more than 100-fold induction in A549 cell and more than 40-fold in HepG2; expression level of the system markedly increased after addition of glycin arm, almost 10-fold enhancement in HepG2 cell. Because basic expression also increased, the induction fold by hypoxia decreased.

双质粒共转染不同细胞株后，在1％O〓环境下培养24h，荧光素酶活性检测结果提示含不同结构域的表达系统均有受缺氧激活的能力，其中GAL-CAD在A549中获得100倍以上的诱导，在HepG2中获得40倍以上的诱导；加入甘氨酸臂后该表达系统的转录活性大为增强，在HepG2细胞中增强了10倍以上，但是由于基础表达也有所增加，受缺氧诱导的倍数下降。

A transposon mutagenesis system applicable for Xanthomonas campestrispv.campestrishas been developed by transferring the transposonTn5gusA5,which contains the gus reporter gene,to the broad host rangvector pLAFR1 and taking the advantage of incompatibility between pLAFR1and pPH1JI plasmids.Five different types of exopolysaccharidedeficient mutants related respectively to eight different mutated sites wereisolated by applying the system to mutagenize the Xcc wild type strain.

本研究将含gus报告基因的转座子Tn5gusA5转座到广谱宿主载体质粒pLAFR1后，利用pLAFR1与pPH1JI质粒的不相容性，建立了一套适合于甘蓝黑腐病菌的转座子诱变体系，应用该体系诱变甘蓝黑腐病菌野生型菌株，成功地分离到5种不同类型、分别涉及8个不同突变位点的胞外多糖突变体。

The carboxyl-terminal fragment of approximately 15ku of zonula occludens toxin gene encoding the 264-399 amino acid residue was amplified by PCR and then cloned into the prokaryotic expression plasmids PBCX.

用PCR扩增了MBP-ZOT质粒上的ZOT基因，该基因编码264～399位氨基酸残基C末端的15ku ΔG片段，并与经改造后的质粒PBCX连接。

In order to elucidate the regulation mechanism of RU5 region to BFV gene expression, BFV3026 provirus DNA was used to construct the plasmids containing different deletion of R region, which were cotransfected with luc report gene locatied behind the IP promoter to BL12 cells, and luciferase activities was assayed, confirming that U5 region could repress the initiation function of LTR as well as IP. The BFV structure genes with different deletion of R region were cloned closely behind to heterogenous CMV promoter, then transfected to 293T cells, RT activity was performed, testifying the R region was required for BFV pol gene expression, and also the function domain was identified within the 100n.t. sequence at the 5′end.

以牛泡沫病毒(Bovine foamy virus， BFV)中国株BFV3026原病毒DNA为材料，构建R区系列缺失质粒，通过对其转染细胞中RT水平及对缺失质粒与luc报告质粒共转染细胞中萤火虫荧光素酶活性的测定，确立U5区对于BFV3026两类启动子LTR和IP均具有负调控作用；同时将带有不同R区的BFV3026结构基因片段克隆于异源启动子CMV之下，通过对其转染细胞293T中RT酶活性的测定，确立R区对于病毒结构基因pol的表达具有一定的调节作用，并将其功能区域初步界定在R区5′端100bp内。

Construction of heavy chain transgenic mice and genome type analysis Linearized transgene plasmids were microinjected into pronuclei of zygotes from CBA×C57BL/6 mice, and transplamted into oviduct of pseudo-pregnant mice.

二、抗体重链转基因小鼠建立及基因型分析将线性化的质粒显微注射入CBA×C57BL/6小鼠的受精卵细胞的雄前核内，然后导入假孕雌鼠的输卵管。

Plasmids carry between 2 and 30 genes.

2在细胞分裂时可见多个。

I have also constructed Kv1.4S229A and Kv1.2S84A mutant plasmids, which can be used for further experiments.

另外我还构建了Kv1.4S229A 和Kv1.2S84A 突变质粒，为下一步的实验奠定了基础。

MAIN OUTCOME MEASURES: Identification of the recombinant plasmids and the expressing mRNA and protein in 293-T cells.

主要观察指标：质粒的酶切鉴定及重组质粒在293-T细胞中mRNA及蛋白水平的表达。

She gently rebuff ed him, but agreed that they could be friends

她婉言拒绝了，但同意作为朋友相处。

If in the penal farm, you were sure to be criticized.

要是在劳改农场，你等着挨绳子吧！