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Firstly, MiCy, Citrine, mKO were inser into expression vectors of pRSETB, and then MiCy, Citrine,mKO were expressed in E.coli and purified by using Ni-NTA collum chromatography.

首先，构建了含有六个组氨酸标签的三种荧光蛋白MiCy、Citrine和mKO，而后在大肠杆菌中表达了MiCy、mKO和Citrine三种蛋白，并用金属鳌合亲和层析进一步纯化，纯化产物使用特异性酶去除His-tag。

Objective: To contructre combinant plasmid pET28a/hES and coexpress human Endostatin and predhPK-5 in E. coli. Methods: mRNA from human liver tissue was exracted, and the Endostatin gene was amplified by RT-PCR.

目的构建人内皮抑素原核表达质粒pET28a/hES，并与简化人纤溶酶原饼环区(predigested human PlasminogenKringle5，predhPK-5)在大肠杆菌中实现共表达。

Methods:The total RNA was extracted from mouse RAW264.7 cells stimulated by LPS.The sequence including the whole length of HMGB1 was amplified by RT-PCR and inserted into pMD-19T.The combinant vector was used as a template for PCR which was cloned into vector pMD-19T,then subcloned into expression vector pET-26b with pelB signal sequence and His-Taq sequence.After transforming E.coli BL21(DE3) and four hours induction by IPTG,HMGB1 expression confirmed by SDS-PAGE and the purification was performed by Ni2+-chelate affinity chromatograph.

脂多糖刺激后的RAW264.7细胞，提取总RNA，经RT-PCR扩增出含HMGB1的目的片段，克隆于pMD-19T载体，再亚克隆至含有pelB引导肽及His-标签肽的高效表达载体pET-26b，转化大肠杆菌BL21（DE3），经IPTG诱导后行SDS-PAGE鉴定目标蛋白表达，用镍鳌合琼脂糖凝胶亲和层析法分离纯化含His-标签肽的目的蛋白。

The combinant vector was used as a template for PCR which was cloned into vector pMD-19T,then subcloned into expression vector pET-26b with pelB signal sequence and His-Taq sequence.After transforming E.coli BL21(DE3) and four hours induction by IPTG,HMGB1 expression confirmed by SDS-PAGE and the purification was performed by Ni2 -chelate affinity chromatograph.

脂多糖刺激后的RAW264.7细胞，提取总RNA，经RT-PCR扩增出含HMGB1的目的片段，克隆于pMD-19T载体，再亚克隆至含有pelB引导肽及His-标签肽的高效表达载体pET-26b，转化大肠杆菌BL21（DE3），经IPTG诱导后行SDS-PAGE鉴定目标蛋白表达，用镍鳌合琼脂糖凝胶亲和层析法分离纯化含His-标签肽的目的蛋白。

AFM images revealed that as the sputtering time was prolonged, the film thickness increased, the film became compacter, and the specific area of the film also increased, thus, the release rate of silver ions increased, which led to improved antibacterial properties. EDX test results indicated that increase in film thickness led to increase in silver weight percentage per unit surface, causing improved antibacterial properties. Moreover, when the film thickness was 1 nm, the inhibition percentage of Staphylococcus aureus and Escherichia coli were 100%.

结果表明：随着薄膜厚度的增大，样品抗菌性能提高；溅射时间延长，薄膜厚度增大，膜层的致密性改善，单位面积上的银含量增加，膜层表面积增大，银离子释放几率增大，是提高抗菌性能的主要原因；当纳米结构银薄膜厚度为1 nm时，对大肠杆菌和金黄色葡萄球菌的抑菌率均达到100%。

Coli. The shuttle co-expression plasmids were introduced into filamentous cyanobacterium-Anabaena sp. PCC 7942 (or 7120) by a triparental conjugative transfer method.

采用双顺反子体系在pTrc载体中成功的实现了A、B和Bc亚基的单独活性表达及A、B亚基共表达。

Owing to growth rapidly and predominance in fecal contaminated water, we could detect and identify culturable E. coli cells in 50 mL of pond water in campus within one working day.

此外，本方法样品需求量低且不需繁复的前处理步骤，分析环境水样中之大肠杆菌，可在一个工作天内得到结果，为大肠杆菌检测提供一个快速、简单及经济的方法。

Escherichia coli O157:H7 was cultured in sterilized fresh water at 4℃; 95~115 days later, plate count and most probable number showed that there was no culturable bacteria.

将大肠杆菌 O157：H7培养于低温贫养条件下，以涂布平板法和最大近似值法检测可培养细菌数，95～ 115d后表明可培养菌数下降为零。

Escherichia coli O157:H7 was cultured in sterilized fresh water at 4℃; 95~115 days later, plate count and most probable number showed that there was no culturable bacteria.

摘 要 将大肠杆菌O157：H7培养于低温贫养条件下，以涂布平板法和最大近似值法检测可培养细菌数，95~115d后表明可培养菌数下降为零。

Escherichia coli O157:H7; viable but non culturable state; acridine orange direct count; direct viable count

H7；活的非可培养状态；吖啶橙荧光显微镜直接计数法；活菌直接镜检计数法

Do you know, i need you to come back

你知道吗，我需要你回来

Yang yinshu、Wang xiangsheng、Li decang,The first discovery of haemaphysalis conicinna.

1〕 杨银书，王祥生，李德昌。安徽省首次发现嗜群血蜱。