利用cDNA-AFLP技术研究副溶血弧菌感染下拟穴青蟹的基因差异表达

The differential gene expression of Scylla paramamosain upon Vibrio parahaemolyticus infection by cDNA-AFLP

研究以实验室分离自汕头牛田洋青蟹养殖区的副溶血弧菌感染的健康拟穴青蟹,利用cDNA-AFLP技术分析感染前后拟穴青蟹肝脏组织基因的转录表达差异,最终获得了23个差异片段,其中20个片段成功测序。BLAST分析表明,序列与已知功能基因具有同源性的有6个,其中5个经real-time PCR重新验证为上调表达,主要涉及参与能量代谢的精氨酸激酶(arginine kinase)和甘油醛3磷酸脱氢酶(glyceraldehyde-3-phosphate dehydrogenase)、磷酸烯醇丙酮酸羧激酶(phosphoenolpyruvate carboxykinase),以及参与转运过程的精氨酰tRNA合成酶(arginyl-tRNA synthetase),同时,还有直接参与免疫防御反应的凝乳状蛋白酶(chymotrypsin-like proteinase)。另外,8个差异片段与已知基因或序列无同源性。

Mud crab Scylla paramamosains is widely cultured in brackish and seawater ponds along the coast of southeast China including Shantou.In recent years,mud crab cultured in Niutianyang reclaiming coastal area of Shantou suffered from serious diseases,especially the bacterial diseases（such as Vibrio parahaemolyticus）,which results in high mortality and great economic loss.In present study,V.parahaemolyticus isolated from Shantou Niutianyang crab culture area was injected to S.paramamosains to study the differential gene expression of the mud crab by cDNA-AFLP technology.Furthermore,20 fragments that exhibited differential expression were discovered and sequenced.The results show that 6 fragments are homologous to the known functional genes,5 of which are up-regulated.They are involved in energy metabolism including arginine kinase,phosphoenolpyruvate-carboxykinase and glyceraldehydes 3 phosphate dehydrogenase,arginyl-tRNA synthetase involved in transport progress,as well as the chymotrypsin-like proteinase involved in defense response.The other 8 fragments were identified to be unknown function genes.The result here shows that the response of S.paramamosain upon V.parahaemolyticus infection is involved in multiple pathways and needs further investigation.