ABSTRACT
Objective To establish a stably overexpressing miR-31 transgenic mouse and detect the expression of miR-31 in the organs and tissues,and to provide qualified tool mice with overexpression of miR-31 in vivo. Methods The miR-31 overexpression vector was constructed by Gateway cloning technology. The vector was injected into fertilized ovum by DNA microinjection technology,then transferred to the pseudopregnant mice and waited for eutocia. Newborn mouse tail DNA was extracted and PCR and agarose gel electrophoresis were performed to identify the positive miR-31 transgenic mice. microRNA was extracted from the organs and tissues of miR-31 transgenic mice and the expression of miR-31 was de-tected by RT-PCR. The expression of Nestin and number of neural stem cells in the nervous system were compared in the positive and WT mice. Results The miR-31 transgenic mice were constructed successfully and bred more than 14 genera-tions in barrier environment. Expression of miR-31 was increased in major organs and tissues. The expression of Nestin and the number of neural stem cells in the positive mice were higher than those in the wild type mice. Conclusions MiR-31 overexpressing transgenic mice are constructed by Gateway cloning technology and the expression of miR-31 is stable in sub-sequent generations. The number of neural stem cells in the nervous system is higher than that in wild-type mice. The miR-31 overexpressing transgenic mice can be a good tool for experimental research of the function of overexpressed miR-31 in vivo and the treatment of nervous system diseases.
ABSTRACT
Studies have shown that some plant-specific NAC (NAM, ATAF1/2, CUC2) transcription factors may increase plants resistance to stress. We screened the genes differentially expressed in transgenic SlNAC1 Arabidopsis compared to the wild type by cDNA microarry, to provide scientific basis for studying the genes related to abiotic stress responses in transgenic Arabidopsis. There were 3 046 genes differentially expressed more than twice in the total 43 604 genes of transgenic SlNAC1 Arabidopsis. Gene ontology analysis was used on genes differentially expressed more than five-fold. Genes relevant to cellular components occupied 33.05%, genes correlated with molecular function accounted for 33.95% and genes pertinent to biological process constituted a 33.00% portion. The genes differentially expressed more than twice were processed through kyoto encyclopedia of genes and genomes pathways enrichment (KEGG) analysis. The total 2 431 genes were involved in 88 different signaling pathways. The screened genes related to abiotic stress responses provide direction and theoretical support for the following research on the downstream genes regulated by NAC and construction of the regulatory networks.