To construct eukaryotic expression vector of mbl gene with codon 54 point mutation , the target sequence in pgem - mbld plasmid , which conains mbl cdna with codon 54 mutant allele , was amplified by pcr . after the cdna fragement and plasmids pci - neo were prepared by digestion with sma i and sal i , the fragment was inserted into sma i and sal i site in pci - neo eukaryotic expression vector , and the recombinant vector , named pci - mbl54 , was obtained . the pci - mbl54 , digested with restriction enzymes , was found to contain the point mutation mbl cdna by agarose gel electrophoresis analysis 本实验以ggc54gacmbl突变为研究对象，选用真核表达质粒pci - neo ，根据已构建好的含54位密码子突变型mbl基因t载体的结构，设计合成新的引物， pcr扩增54位密码突变型mbl基因，凝胶回收，双酶切pcr产物和pci - neo质粒， t4连接酶连接，将前者克隆至后者的sma和sal位点，转化大肠杆菌xl - 1blue ，氨苄选择培养。
In order to investigate the role of mannose receptor ( mr ) of human sperm , the zona free hamster eggs were pre - incubated with purified mr ( pmr ) isolated from motile human sperm by mannose - agarose gel affinity chromatography . the ultrastuctural alteration and cortical granule exocytosis of the eggs were then observed by transmissian electron microscope and tritc - lca immunofluorescence microscope , respectively . the mice were immunized with pmr and the antiserum was raised . after capacitation and induction of the acrosome reaction , the human spermatozoa and oocytes were incubated with the antiserum . then the sperm penetration assay was undertaken 为了进一步探讨人精于mr在精卵融合中的作用，本文采用改良后的甘露糖-琼脂糖凝胶亲和层析法分离纯化人精子mr ，并将提纯的人精子甘露糖受体( purifiedmannosereceptor , pmr )作用于去透明带的金黄地鼠卵母细胞，运用透射电子显微镜技术和罗丹明偶联的兵豆凝集素( tritc - lca )免疫荧光标记技术观察pmr对卵子的影响。
Thermocycled pcr samples were resolved electrophoretically on 1 . 5 % agarose gels and taken photos using a polaroid camera . the statistical results were analyzed by the spss software , and the cluster figures were obtained . conclusions could be drawn from the study as following : 1 ) the molecular systematics of 57 species of crickets , which belong to 26 genera 7 family in grylloidea , had been studied by the approach of rapd 本项研究在依据外部形态分类鉴定及前人工作的基础上，采用rapd技术，通过对蟋蟀总科7科26属57种蟋蟀基因组dna的rapd图谱的比较研究，在分子水平上探讨这些类群的分类地位和亲缘关系，为丰富蟋蟀总科的分子系统学研究，并为进一步完善蟋蟀总科的分类系统，揭示其系统发育及演化提供分子水平的依据。
Main works and their important results are showed as following . firstly , the cdna encoding bullfrog gh is amplified by use of the total rna , extracted from adult bullfrog ' s pituitary , as the template , pi ( 5 ' - cggatcc atg gct tca ggg tta ggc ) and p2 ( 5 ' - cgaattc tta aaa ggt gca gtt gct ) as the primer pair and one particular cdna product , 660bp , was obtained after the rt - pcr - the product was purified by using dna agarose gel extraction method . after the purified cdna and pmd18 - t vector were ligated at 16 over night , the ligation products were transformed into e . coli strain dh5a and then the transformants were screened by clone pcr method 首先，以成体牛蛙的脑垂体总rna为模板，进行rt ? pcr扩增得到约660bp的特异性pcr产物带，切下琼脂糖凝胶中的特异产物带进行cdna胶回收，将cdna与pmd18 ? t载体进行t ? a克隆连接并转化到dh5 e . coli后，进行菌落pcr 、质粒酶切鉴定，筛选出阳性菌株，测序结果经blast分析，与已报道的牛蛙生长激素基因高度同源，证实此阳性克隆为牛蛙生长激素基因转化子，命名为pbfgh 。
Materials and methods in our study , first the e . coli bl21 transformed already are cultivated in smaller scale and then the plasmids dna were extracted by the methods of alkaline lysis . the plasmids dna extraction were processed 37 overnight by the restrictive endo - incisase bam h i and xho i . the incision products were used to check the integrality and variability of the recombinant plasmids dna by 1 % agarose gel elec - trophoresis 同预想结果基本吻合，大规模培养后纯化得到的融合蛋白通过sds page显示在52kda处有一条带，而酶切后产物电泳结果显示在26kda处分另有两条蛋白带，其中一条为gst ，另一条为hbrp ，基本符合实验结果； hbrp对tpk活性的抑制呈剂量依赖性。
Methods : the effects of different neurotrophic factors on the growth and differentiation of neural stem cells were observed by cells counting and immunofluorescence staining . the levels of rara mrna and rxra mrna in differentiated neural stem cells were assayed by rt - pcr . agarose gel electrophoresis and image analysis 方法应用细胞计数和免疫荧光细胞化学法，研究不同神经营养因子对神经干细胞增殖及分化的影响；应用rt - pcr 、琼脂糖凝胶电泳和紫外分光图象分析法检测神经干细胞分化过程中rar和rxr mrna表达量的改变。结果1
In order to detect the effect of human sperm mannose - ligand receptor on the fertilization ability , in the study reported here mannose - ligand receptors ( mrs ) were purified from human sperm by modified mannose - agarose gel affinity chromatography coloumn and determined protien concentration by lowry , preincubated zona - free hamster oocytes with four purified mannose - ligand receptor ( pmr ) concentrations before sperm penetration assay ( spa ) to test the pmrs cell biology nature of inhibition to fertilization 本研究用改良后的亲和层析法分离纯化mr ， lowry法测定其蛋白质浓度，在精子穿透试验( spermpenetrationassay ， spa )模型中定量研究其对精卵融合能力的影响并检测其细胞生物学活性；以已知浓度的pmr ( purifiedmannose - ligandreceptor )干预精子半透明带试验，观察用pmr预处理半透明带对精子与透明带结合的影响。
It has marked apoptotic induction on mouse spleen cells tested by dna agarose gel electrophoresis analysis and flow cytometric assay in certain degree . it also can decrease the activity of lactate dehydrogenase ( ldh ) outside of mouse the spleen cells and increase the quantity of reduced gsh inside the mouse spleen cells 而且一定浓度范围内可引起小鼠脾淋巴细胞dna发生特征性降解，出现dnaladders ，引起染毒组细胞凋亡率显著性升高，但细胞凋亡率与剂量不呈直线相关关系。
