ABSTRACT
The aim of this study is to clone the mouse T-bet promoter and enhancer, construct and identify the firefly luciferase reporter gene plasmid pGL4.10-TBX21pr-CNS for T-bet transcription regulation study and its function in signaling of multiple sclerosis. The promoter and CNS of T-bet were predicted by bioinformatics assay. The predicted fragment of mouse T-bet promoter plus CNS was amplified by PCR and cloned into pGL4.10. The recombinant plasmid pGL4.10-TBX21pr-CNS was transferred into Escherichia coli DH5α. The positive clone was identified by double digestion with Kpn I and Sfi I and DNA sequencing. Finally, pGL4.10-TBX21pr-CNS was cotransfected with pRL-TK into 293T cells and Jurkat cells, pRL-TK and pGL4.10 as a control. The luciferase activity in 293T cells (P = 0.012 2) and Jurkat cells (P = 0.002 2) was higher than that of the control group. A fragment of 1 028 bp mouse T-bet promoter plus 1 308 bp CNS was successfully cloned and the firefly luciferase reporter gene plasmid pGL4.10-TBX21pr-CNS was constructed. In 293T cells and Jurkat cells, pGL4.10-TBX21pr-CNS has the promoter functions. This work offers a basic material for the research of T-bet transcription.
Subject(s)
Animals , Mice , Gene Expression Regulation , Genes, Reporter , Genetic Vectors , Luciferases , Multiple Sclerosis , Plasmids , Promoter Regions, Genetic , T-Box Domain Proteins , GeneticsABSTRACT
A field investigation was conducted on the floral syndrome and breeding system of Corydalis edulis located in natural populations in campus of Anhui Normal University by out-crossing index, pollen-ovule ratio, artificial pollination and bagging experiment. The results showed that the plant was in bloom from March to May and flowering span among populations was 72 days. The flowering span for a raceme was 14-24 days. The life span of one single flower was approximately 5-10 days. Spatial positioning of stigma and anthers were spatially desperation on the day of anthesis. The filaments were shorter than the styles through pollen vitality and stigma receptivity experiments. A self-pollination breeding system was reflected by OCI 3, pollinators were required sometimes; A complex cross bred was indicated by P/O = 857.14, combined with the results of the bagging and artificial pollination experiment, the breeding system of C. edulis was mixed with self-pollination and outcrossing. The special floral structure and pests destroying may have a certain impact on seed-set rate.
Subject(s)
Breeding , Corydalis , Physiology , Flowers , Physiology , Pollination , Physiology , Reproduction , PhysiologyABSTRACT
<p><b>OBJECTIVE</b>To reveal the flowering characteristics and provide theoretical basis for the breeding and cultivation of Lamium barbatum.</p><p><b>METHOD</b>The field observation and investigation of blooming phenophase were applied in the study growth dynamics of inflorescence and the period of the floret were measured. Then the statistic analyses of them were carried out. The pollen viability was evaluated by TTC test and the stigma receptivity was evaluated by benzidine-H2O2 method.</p><p><b>RESULT AND CONCLUSION</b>The flowers of L. barbatum were bisexual, with 4 to 14 flowers in an inflorescence. The population came into bloom from April to June and the blossom of a single flower could last about 3 days. Pollination peak lasted 8 hours from the very beginning of blooming, and the high stigma receptive stage remained during the same period of time. The results indicated that the breeding system of L. barbatum was outcrossing with self-compatibility and pollinators were needed based on the outcrossing index. The most frequent floral visitors were bees. The suitable breeding strategy was also discussed.</p>