RESUMEN
Cynanchum auriculatum is a traditional herbal medicine in China and can grow in saline soils. However, little is known in relation to the underlying molecular mechanisms. In the present study, C. auriculatum seedlings were exposed to 3.75 and 7.5 salinity. Next, transcriptome profiles of leaves were compared. Transcriptome sequencing showed 35,593 and 58,046 differentially expressed genes (DEGs) in treatments with 3.75 and 7.5, compared with the control, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of these DEGs enriched various defense-related biological pathways, including ROS scavenging, ion transportation, lipid metabolism and plant hormone signaling. Further analyses suggested that C. auriculatum up-regulated Na+/H+ exchanger and V-type proton ATPase to avoid accumulation of Na+. The flavonoid and phenylpropanoids biosynthesis pathways were activated, which might increase antioxidant capacity in response to saline stress. The auxin and ethylene signaling pathways were upregulated in response to saline treatments, both of which are important plant hormones. Overall, these results raised new insights to further investigate molecular mechanisms underlying resistance of C. auriculatum to saline stress.
Asunto(s)
Perfilación de la Expresión Génica , Hojas de la Planta/genética , Estrés Salino/genética , Vincetoxicum/genética , Antioxidantes/metabolismo , Ontología de Genes , Anotación de Secuencia Molecular , Análisis de Secuencia , Vincetoxicum/metabolismoRESUMEN
<p><b>OBJECTIVE</b>To investigate the anti-tumor activity of CCL21-exCD40L eukaryotic expression vector.</p><p><b>METHODS</b>CCL21-exCD40L fusion gene were constructed by overlap PCR connecting CCL21 and exCD40L through a flexible linker (Gly3Ser)4, and then was cloned into expression vector pcDNA3.1(+). pcDNA3.1(+)/CCL21 and pcDNA3.1(+)/exCD were constructed as negative control. Wsestern blot was used to identify the fusion protein. CHO cells was transfected with pcDNA3.1(+)/CCL21-exCD, pcDNA3.1(+)/CCL21 and pcDNA3.1(+), respectively. The chemotatic function of the expressed product was detected by Transwell method and its anti-tumor activity was tested with vivo transfection.</p><p><b>RESULTS</b>Gene sequencing and restrictive digestion proved the successful construction of pcDNA3.1(+)/CCL21-exCD40L,and its expression was conformed by western blot. The transfectant supernantes of pcDNA3.1(+)/CCL21-exCD40 group had a significant chmotactic function to DCs, of which the cell numbers passing through the film was 14.95 times of blank control every high power microscope visual field. After tumor orthotoic injection of plasmid carrying fusion gene in Balb/c mouse, the tumor mass reduced remarkablely, and all the mouse in fusion gene group survived after 4 weeks.</p><p><b>CONCLUSION</b>CCL21-exCD40L fusion protein had a remarkable function to DCs and it can inhibit tumor growth and prolong the mouse survival time, which is more effective than all control group.</p>
Asunto(s)
Animales , Ratones , Ligando de CD40 , Genética , Farmacología , Células CHO , Línea Celular Tumoral , Quimiocina CCL21 , Genética , Farmacología , Neoplasias del Colon , Terapéutica , Cricetulus , Células Dendríticas , Fisiología , Terapia Genética , Ratones Endogámicos BALB C , Proteínas Recombinantes de Fusión , FarmacologíaRESUMEN
Mucosal adjuvants play important roles in vaccine development. By now, the common used mucosal adjuvants can be divided into three categories: the bacterial derivatives, cytokines and chemokines, and antigen delivery systems. Progresses of the three kinds of adjuvants were reviewed to give a reference to novel vaccine research.