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AIM: To prepare gfp-bcl-XL-contained recombinant adenovirus(rAd-gfp-bcl-XL).METHODS: Bcl-XL gene was amplified from pEGFP-C3-bcl-XL, subcloned into shuttle plasmid and formed transfer plasmid of pAdTrack-CMV-bcl-XL. Then pAdTrack-CMV-bcl-XL was linealinzed with PmeI and co-transformed into BJ5183 bacteria with adenovirus genomic plasmid of pAdEasy-1. The identified recombinant adenovirus plasmid was digested with PacI and transfected into 293 cells to package recombinant adenovirus particles. The target gene was detected by PCR.RESULTS: There were about 35% positive recombinant bacterial clones after the co-transformation of pAdTrack-CMV-bcl-XL and pAdEasy-1 into BJ5183. Recombinant adenovirus particle were produced and further amplified after the transfection of pAdEasy-1-gfp-bcl-XL into 293 cells. PCR test indicated that the recombinant Ad contained bcl-XL gene. The titer of the purified rAd-gfp-bcl-XL was 6.5×1012 PFU/L. CONCLUSIONS: The homologous recombination in bacteria is a convenient and high efficient method to prepare rAd-gfp-bcl-XL. This affords a good gene transfer vector for the gene therapy in human’s diseases. 相似文献
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Yasuhiro Tomitaka Tohru Yamashita Kazusato Ohshima 《Journal of General Plant Pathology》2007,73(3):197-208
The genetic structure of the populations of Turnip mosaic virus in Kyushu and central Honshu, Japan was assessed. The host specificity of isolates was determined, and their gene sequences
compared utilizing a population genetic approach. Phylogenetic analysis of partial sequences revealed that 32 of 49 Honshu
isolates (65%) collected during 1997–2001 belonged to the basal-BR group as did 23 of 64 isolates from Kyushu. All these basal-BR
isolates infected both Brassica and Raphanus plants. However, analyses of the positions of recombination sites in five regions of the genome (one third of the full sequence)
showed that at least four intra-lineage recombinants were present in these populations. These analyses showed that Kyushu
and Honshu shared none of these subpopulations, and genetically distinct basal-BR populations were present in the two districts.
We conclude that different basal-BR subpopulations had expanded into those districts.
The nucleotide sequences are deposited in the DDBJ/EMBL/GenBank databases under accession numbers AB267281-AB267376. 相似文献
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Pure lines derived from multiple parents provide abundant variation for genetic study.However,efficient genetic analysis methods and user-friendly software are still lacking.In this study, we developed linkage analysis methods and integrated analysis software for pure-line populations derived from four-way and eight-way crosses.First, polymorphic markers are classified into different categories according to the number of identifiable alleles in the inbred parents.Expected genotypic probability is then derived for each pair of complete markers, and based on them a maximum likelihood estimate(MLE) of recombination frequency is calculated.An EM algorithm is proposed for calculating recombination frequencies in scenarios that at least one marker is incomplete.A linkage map can thus be constructed using estimated recombination frequencies.We describe a software package called GAPL for recombination frequency estimation and linkage map construction in multi-parental pure-line populations.Both simulation studies and results from a reported four-way cross recombinant inbred line population demonstrate that the proposed method and software can build more accurate linkage maps in shorter times than other published software packages.The GAPL software is freely available from www.isbreeding.net and can also be used for QTL mapping in multi-parental populations. 相似文献
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家蚕标记基因位点与数量性状基因位点重组参数的最大似然估计法 总被引:8,自引:2,他引:6
利用分离世代群体(BC_1或F_2),估算家蚕标记基因位点与数量性状基因位点重组参数的最大似然估计法,求解似然方程组的EM迭代算法。通过120次计算机模拟和一个生产实例验证了方法的可行性。 相似文献
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Sopid Sawangjit Orawan Chatchawankanphanich Pissawan Chiemsombat Tipvadee Attathom James Dale Supat Attathom 《Journal of General Plant Pathology》2005,71(4):314-318
Nucleotide sequences of the three distinct Tomato yellow leaf curl Thailand virus (TYLCTHV) strains (CM, NK, SK) were analyzed for recombination events. Recombination detection program analyses and a sequence alignment survey provided evidence of recombination between AC1 sequences of TYLCV, TYLCTHV-[MM], and TYLCTHV-[NK] as major parents and of ToLCLV, ToLCTWV, and TYLCTHV-[SK] as minor parents of TYLCTHV-[NK], -[SK], -[CM], respectively. The results further support the notion that interspecies recombination may play a significant role in geminivirus diversity and their emergence as important pathogens. 相似文献
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Poss M Ross H Rodrigo A Terwee J Vandewoude S Biek R 《Veterinary immunology and immunopathology》2008,123(1-2):154-158
Feline immunodeficiency virus (FIV) is a lentivirus that has been identified in many members of the family Felidae but domestic cats are the only FIV host in which infection results in disease. We studied FIVpco infection of cougars (Puma concolor) as a model for asymptomatic lentivirus infections to understand the mechanisms of host–virus coexistence. Several natural cougar populations were evaluated to determine if there are any consequences of FIVpco infection on cougar fecundity, survival, or susceptibility to other infections. We have sequenced full-length viral genomes and conducted a detailed analysis of viral molecular evolution on these sequences and on genome fragments of serially sampled animals to determine the evolutionary forces experienced by this virus in cougars. In addition, we have evaluated the molecular genetics of FIVpco in a new host, domestic cats, to determine the evolutionary consequences to a host-adapted virus associated with cross-species infection. Our results indicate that there are no significant differences in survival, fecundity or susceptibility to other infections between FIVpco-infected and uninfected cougars. The molecular evolution of FIVpco is characterized by a slower evolutionary rate and an absence of positive selection, but also by proviral and plasma viral loads comparable to those of epidemic lentiviruses such as HIV-1 or FIVfca. Evolutionary and recombination rates and selection profiles change significantly when FIVpco replicates in a new host. 相似文献
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转病毒移动蛋白及复制酶基因烟草的协生和重组风险分析 总被引:2,自引:0,他引:2
对转化番茄花叶病毒 (ToMV)、烟草花叶病毒 (TMV)移动蛋白 (MP)基因和黄瓜花叶病毒P1株系(CMV P1)部分复制酶基因的 3种转基因烟草 ,分别接种马铃薯X病毒 (PVX)、马铃薯Y病毒 (PVY)、CMVRB株系 (CMV RB)和TMV。症状观察和ELISA测定发现 ,各病毒在转基因烟草和非转基因烟草上的症状及病毒浓度没有显著差异 ,表明这些病毒在转基因烟草上没有协生作用发生。对转化ToMVMP基因和转化CMV P1部分复制酶基因的转基因烟草 ,分别接种TMV和CMV RB ,定期进行生物学、ELISA和免疫捕获反转录PCR(IC RT PCR)检测 ,在 16个月的连续测定中未发现病毒重组现象。 相似文献