dna测序

• Compared with the results of dma direct sequencing , pcr - sscp has the same results as that except for failing to check out the rpob gene mutation of a rifampin - resistant isolate . it proves that pcr - sscp is a rapid and sensitive mutation testing method
  与dna测序结果相比较， pcr - sscp除一个耐药株的rpob基因突变没有检出外，其它检测结果均与dna测序结果相一致，这进一步说明pcr - sscp是一种快速、灵敏的突变检测技术。
• Conclusion : by restriction enzyme secting , ligating , transforming , restriction enzyme analysis , and final dna sequencing , the pbd - i and pbd - ii gene were proved to be recombinated with the expression vector and the recombinated vector ppd - 1 and ppd2 were transformed successfully
  结论：经过酶切、连结，构建成重组质粒ppd上、 ppd上，再经转化、抽提质粒及酶切分析，最后经dna测序证实， rticr扩增的pbd i 、 pbd 11基因与piflpdt ” xsi表达载体构建成功。
• Part ii screening of tnfa mimotopes from phage display peptide library : based on the results of screening tnfa binding - peptides , we have tried to use neutral tnfa mcab j1d9 as target to screen tnfa mimotopes from c7c phage display peptide library , which may be another form of antagonist for tnfa , and the mimotopes were identified by sandwich elisa . after 3 rounds of screening , we got 9 phage clones identified as positive clones which can bind with mcab j1d9 . we also identified the binding between mimotopes and tnf receptor by competitive elisa , and the results showed strongly binding . the amino acid sequence results shown three different sequences : c - rrpaqsg - c - nkhnrki - c and c - rgmsrki - c
  在对噬菌体环七肽库进行三轮亲和性筛选后，随机挑选20个噬菌体克隆， elisa鉴定出9个阳性克隆，经dna测序推出三种氨基酸序列： c - rrpaqsg - c 、 c - nkhnrki - c和c - rgmsrki - c ，其中优势克隆序列为c - rrpaqsg - c ；鉴定结果显示阳性克隆能够与tnf受体结合，并且能够阻断tnf与受体的结合，提示筛选得到的环七肽克隆展示肽具有tnf的抗原性及与tnf受体结第一军医大学顾士学位论文合的特性，为tnfa表位模拟肽。
• 2 heterogeneity and evolution of 5s rdna pattern of intragenomic and interspecific variation of 5s rdna hi five pines ( including subgenus strobus and pinus ) were studied through cloning and sequencing multiple 5s rdna copies the length of 5s rdna unit is 658 - 728 bp hi diploxylon pines while 499 - 521 bp hi the haploxylon p . bungeana
  Ssif7 ： na的序列变异与分子进化利用分子克隆和dna测序分析了油松、云南松、马尾松、白皮松和不同遗传背景的高山松居群的ssilna基因序列变异及基因进化规律，得到以下主要结果： （ 1 ） ssrdna的结构特征。
• The amplification system was optimized so that the pcr with different primers can be carried out under the same condition . the pcr products were sequenced using sanger ' s terminator and fluorescent label techniques . sequences were analyzed and compared base on sequencing analysis3 . 4 and seq / ede software
  方法根据mtdna控制区及其周围区域的序列，设计多对引物，探索优化扩增体系，使扩增条件能够同时满足多对引物的需要，用sanger末端终止法及荧光标记技术对样本进行dna测序， sequencinganalysis3 . 4和seq ede软件进行序列分析和比对。
• Results : designed and synthesized the coding gene of echistatin . and constructe its single copy expression vector , identify the vector by dna sequencing . but it could not express echistatin in e . coli . designed and synthesized the modified coding dna of echistatin , and constructe its two copy expression vector , identify the vector by dna sequencing , but it could not express echistatin in e . coli
  结果设计并合成了echistatin编码基因dna序列，构建了echistatin单拷贝表达载体，经dna测序鉴定正确后， sds - page及western - blotting结果表明： echistatin在上述菌株中未获得高效表达。
• [ methods ] by using the overall rna of our previously cultured human melanoma cell line ( a375 ) , full length fasl gene is detected by rt - pcr . using the cdna as template , . the extracellular domain of fasl ( fasl - ecd , 127 - 278aa ) is amplified by pcr . the pcr products are directly cloned into t vector pmd - 18t
  L方法采用新鲜人黑色素瘤细胞( a375 ) ，抽提该细胞的总rna ，进行rt一pcr反应分析a375内fasl全长编码基因的转录表达，以a375细胞cdna为模板，用pcr产物直接克隆法扩增人fasl一ecd (人fasl胞外区)的编码基因，即127一278位氨基酸残基，而后将pcr产物直接克隆于pmd一1st载体中，获得重组质粒pmd一t - fasl一ecd ，进行dna测序。
• The e2 genes above of the prevalent strain ( guangxi yulin strain ) were cloned respectively into secreted expression vector ppic9k of eukaryotic expression system p . pastoris and transformed into p . pastoris by electroporation after linearization , 25 high - copied transformants were obtained by g418 screening . it was proved that the e2 genes were integrated stably into chromosome of p . pastoris by dot blot and dna sequencing
  猪瘟病毒e2基因的真核表达：分别将csfv两个代表株的e2基因克隆入毕赤酵母( p . pastoris )分泌型表达载体ppic9k中，酶切线型化后电穿孔导入p . pastotis进行整合，经g418筛选得到25个高拷贝转化子，经dna斑点试验和dna测序证明外源基因e2稳定地整合到p . pastoris染色体中。
• By elisa analysis , inhibition of binding of clq with the c ! q receptors on u937 cells and competitive inhibition of binding of clq with aggregated immunoglobulin g b y selected phage clones , and dna sequencing , a number of similar , but not identical , sets of sequences of clq - binding clones were identified . the deduced amino acid sequences of selected 9 peptides are wyegpftlytwp , hwdpfslsayfp , ltqhnspffllp , tsnpfflwypqp , qtpfqlw , npfnwts , spfxlts , fltwldp and fstflyp . they show significant efficiency to inhibit the binding of clq with the clq receptors on u937 cells and / or aggregated immunoglobulin g , which suggest that the selected peptides contain the modeling epitopes of clq receptor to bind the collage - like region or igg to bind the head domain of clq
  然后，采用噬菌体肽库技术，以c1q为钓饵蛋白，从12肽库和环7肽库中亲和筛选能与c1q结合的噬菌体克隆，经elisa 、 u937细胞配体结合抑制试验、 aigg竞争抑制试验及dna测序，获得了9个具c1q抑制活性克隆的dna序列，其相应的氨基酸序列为： wyegpftlytwp 、 hwdpfslsayfp 、 ltqhnspffllp 、 tsnpfflwypqp 、 qtpfqlw 、 npfnwts 、 spfxlts 、 fltwldp 、 fstflyp ，它们可能模拟c1qr和或igg的c1q结合表位并抑制c1q的活性。
• First , a pair of pcr primers was designed to isolate fmdv - vp1 gene according to the published fmdv - vp1 sequence . after pcr of dna isolated from the tissues , the fmdv - vp1 gene was cloned into cloning vector . positive clones were analysed with restriction enzyme digestions and further identified with sequence analysis
  首先，根据已知的fmdv基因序列，设计特异性引物，扩增出fmdv - vp1基因，将克隆到的vp1基因连接到测序载体上后用dna测序仪对该序列进行测序。