This procedure was simple , time - saving , highly sensitive and reproducible . based on the improved procedure , nearly 16 , 000 cdna fragments were analyzed between the immature siliques of the wide type and ast mutant , and 28 differential cdna fragments were screened 采用这个改良的的方法，分析了拟南芥野生型和ast突变型植株未成熟角果中16 ， 000个cdna扩增产物条带，从中筛选出28个差异条带。
Then , there are simulations of an example to test the way presented . at last , there is a simple introduction of active fault - tolerant control . the results of the simulations indicate the validity of the fault diagnosis arithmetic based on neural network observer 最后的仿真结果表明,对于模型未知的非线性系统,本文所提出的基于神经网络的故障诊断方法有效,对于系统可能出现的突变型和缓变型故障均能得到很好的诊断结果,并具有一定的鲁棒性。
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。
The plasmids pci - mbl54 containing full length of mutations mbl cdna were propagated hi escherichia coli xl - 1 blue , then the extracted and purified pci - mbl54 were used to transfect dhfr ( - ) chinese - hamster ovary ( cho ) cells . after screeened with g418 and cloned , 4 g418 - resistent clones were randomly selected for detection of mrna expression by rt - pcr and molecular beacons . it was found that all of the 4 positive cell clones expresse mbl analogue as detected in transcription level 抽提、鉴定、纯化重组质粒后，脂质体转染法将重组质粒导入中国仓鼠卵巢细胞( cho - dhfr ~ - )中， g418选择转染子并克隆化培养，经rt - pcr和分子灯塔探针杂交鉴定其mrna转录，获得4株稳定表达54位密码突变型mbl的cho细胞。
