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Insect resistance to Bt toxin is related with the variation of cadherin-like protein gene.cDNA fragment of cadherin-like protein gene in the fifth instar larva of oriental tobacco budworm was amplified using degeneracy primers and RT-PCR technique,and the target fragment was further sequenced after being cloned into pMD18-T vector plasmid.

本研究以烟夜蛾五龄幼虫中肠的总RNA为模板，根据已经克隆的其他昆虫的类钙粘蛋白基因序列设计引物，利用RT-PCR技术扩增出了烟夜蛾类钙粘蛋白基因的cDNA片段，将其连接到pMD18-T载体，转化大肠杆菌后筛选阳性克隆并进行序列测定，测序得到的1 335 bp的片断编码444个氨基酸残基。

With primers designed according to sequence(AY052417) of diapause pupae of oriental tobacco budworm registered on GenBank, a double-strand cDNA fragment of DH gene was obtained from RT-PCR amplification and was further cloned into pGEM-T Easy vector.

根据GenBank发表序列(AY052417)设计引物，以烟夜蛾(Helicoverpa assulta Guenée)滞育蛹为材料，利用RT-PCR技术扩增到滞育激素(diapause hormone，DH)cDNA，并通过TA克隆的方法将其克隆至pGEM-T Easy载体上，对目的基因进行测序，并利用设计的酶切位点将目的基因从T载体上切下。

There is neither paper about detecting DsMV by DBH or IC-RT-PCR nor report about comparing ELISA with these methods published before. Therefore, the DsMV detection methods developed in this study can be applied to virus certification scheme of calla lily and help to produce healthy seedlings in Taiwan.

之前未曾有以DBH或IC-RT-PCR检测DsMV之报告，或这些方法与ELISA比较的相关报导，因此希望本研究所研发的DsMV检测方法未来可应用於每芋种苗病毒验证作业，以协助国内业者生产健康海芋种苗。

METHODS:The expression of Ki67 in three breast caner cell lines was detected by half-quantitative RT-PCR and Western blot assay.

采用半定量RT-PCR和Western blot分别检测3株乳腺癌细胞中Ki67mRNA和蛋白表达水平。

The effect of carotenoid on the bcl-2 gene expression of breast cancer cell lines MCF-7 cell was determined by the method of RT-PCR.

采用RT-PCR方法检测类胡萝卜素对MCF-7细胞株的Bcl-2基因表达的影响，均呈不同程度的下调。

Methods The target DNA sequence of yrdC was obtained from human spleen tissue by using RT-PCR,construct high titer recombinant adenovirus Ad-yrdC by using AdEasy adenovirus carrier system,detect yrdC protein expression by using Western blot and immunohistochemical method,set up control group and transfected Ad-Null group,and study the effects Ad-yrdC on BGC-823 cell line by drawing cell growth curve and MTT chromatometry.

方法用RT-PCR方法获取人脾脏组织yrdC基因序列，采用AdEasy腺病毒载体系统构建携带yrdC基因的高滴度重组腺病毒Ad-yrdC，将其转染胃癌BGC-823细胞，应用Western blot和免疫组织化学法检测yrdC蛋白表达，设对照组和转染Ad-Null组，并分别绘制细胞生长曲线和MTT比色法研究Ad-yrdC对BGC-823细胞的影响。

The chondrocytes of different generation were observed with light-microscope and transmission electron microscope for cellular growth and ultromicrostructure, with the method of MTT assay for grow curve and proliferation, with alcian blue test for GAG of ECM, with immuocytochemistry and RT-PCR (reverse transcript -polymerase chain reaction)for type Ⅱcollagen, with flow cytometry for cell life cycle ,histochemistry for S-A-β-galand so on by which to identify cataplasia and senescence of chondrocytes cultured in vitro.

对软骨细胞退变老化进行观察，采用相差显微镜观察其生长情况，透射电镜观察细胞结构，阿力新蓝染色检测胞外基质硫酸GAG含量和结构，免疫细胞化学法和RT-PCR方法检测Ⅱ型胶原鉴定软骨细胞，以MTT比色法描绘生长曲线、检测生长状态，组化法检测老化相关β-半乳糖苷酶，流式细胞仪分析细胞周期和增殖指数。3。

The third passage chondrocytes were divided into blank group, different desity PAP groups, different desity glucosaminsalfate groups which were passaged to 4th generation and contrast to the 2nd passage group. The chondrocytes of different groups were detected with the method of histochemistry for S-A-β-gal,and with alcian blue test for the content and constructure of GAG of ECM, immuocytochemistry for type Ⅱcollagen and PCNA, MTT assay for proliferation, RT-PCR for type Ⅱcollagen and Aggrecan, flow cytometry for cell life cycle and proliferation index,by which to observe PAP's function regarding to the appearance and functional status in the process of chondrocyte's cataplasia and senescence.

将P3软骨细胞分为空白对照组、鹿茸多肽不同浓度组、硫酸氨基葡萄糖不同浓度组进行传代培养，同时以P2代软骨细胞为对照组，进行组化检测老化相关β-半乳糖苷酶，阿力新蓝染色检测胞外基质硫酸GAG含量和结构，MTT比色检测增殖，免疫细胞化检测PCNA和Ⅱ型胶原，RT-PCR检测Ⅱ型胶原、Aggrecan蛋白，流式细胞仪分析细胞周期和增殖指数等方法，对鹿茸多肽抗软骨细胞退变老化进行分子生物学研究。4。

The 3rdpassage chondrocytes were divided into blank group, different concentration PAP groups,different concentration glucosaminsalfate groups and were sequently passaged to 4thgeneration. The 2nd passage chondrocytes was contrasted as young cells group. Thechondrocytes of different groups were detected with the methods of histochemistry forS-A-β-gal, and with alcian blue test for the content and constructure of GAG of ECM,immuocytochemistry for typeⅡcollagen and PCNA, MTT assay for proliferation, RT-PCRfor typeⅡcollagen and Aggrecan, flow cytometry for cell life cycle and proliferationindex,by which to observe PAP"s function regarding to the appearance and functional status inthe process of chondrocyte"s cataplasia and senescence.

将P3软骨细胞分为空白对照组、鹿茸多肽不同浓度组、硫酸氨基葡萄糖不同浓度组进行传代培养，同时以P2代软骨细胞为对照组，进行组化检测老化相关β-半乳糖苷酶，阿力新蓝染色检测胞外基质硫酸GAG含量和结构，MTT比色检测增殖，免疫细胞化检测PCNA和Ⅱ型胶原，RT-PCR检测Ⅱ型胶原、Aggrecan蛋白，流式细胞仪分析细胞周期和增殖指数等方法，对鹿茸多肽抗软骨细胞退变老化进行分子生物学研究。4。

Total RNA was extracted from flowers of cattleya, according to the conserved acid sequence for ACC oxidase in other orchids, we designed a pair of oligo nucleotide primers. Using RT-PCR method, a cDNA fragment about 967 base pair, which encoded 321 predicted amino acid residues, was amplified.

以卡特兰花瓣为试材，提取其总RNA，并根据其他兰花的ACC氧化酶(1-aminocyclopropane-1-carboxylic acid oxidase， ACO)基因保守序列设计一对特异性引物，通过RT-PCR法克隆得到1条967bp的卡特兰ACO cDNA片断，共编码321个氨基酸残基。

I am accused of being overreligious," she said in her quiet, frank manner,"but that does not prevent me thinking the children very cruel who obstinately commit such suicide.""

客人们在卡罗利娜·埃凯家里，举止就文雅一些，因为卡罗利娜的母亲治家很严厉。

Designed by French fashion house Herm è s, this elegant uniform was manufactured in our home, Hong Kong, and was the first without a hat.

由著名品牌 Herm è s 设计，这件高贵的制服是香港本土制造，是我们第一套不配帽子的制服。