实时荧光定量PCR检测球孢白僵菌热休克蛋白基因hsp70在几种胁迫条件下的表达

本研究运用Realtime-PCR技术,对球孢白僵菌Beauveria bassiana hsp70基因在不同胁迫条件下的表达情况进行了检测。从mRNA转录水平探讨了不同胁迫条件对球孢白僵菌hsp70基因表达的影响。结果表明:38℃高温胁迫下,30min时表达量达到最高峰,为对照样品的10.18倍。随后表达量开始下降,至180min时,其表达量降为最低,为对照样品的2.85倍;4℃低温胁迫下,2h检测到hsp70的表达量下降至最低点,为对照样品的0.25倍。随后表达量开始回升,至10h表达量始终维持在对照样品的1.4-1.5倍左右;紫外胁迫下(波长253.7nm),3min后hsp70的表达量快速上升至最高峰,为对照样品的2.33倍。随后表达量迅速下降,至60min表达量始终维持在对照样品的0.2倍左右。因此推测,hsp70基因在球孢白僵菌抵抗高温、低温和紫外三种胁迫方面都可能具有重要作用。同时研究结果也表明,球孢白僵菌hsp70基因启动子在逆境下可引导基因高效表达,因而在抗逆工程菌株构建方面可能具有重要的应用价值。

Expression analysis of hsp70 gene from Beauveria bassiana under several stress conditions by Realtime-PCR

The expression of hsp70 gene from Beauveria bassiana under several stress conditions was detected using Realtime-PCR, and the effects of different stress conditions on expression of hsp70 gene on mRNA level were explored. The results showed that under high temperature stress (38℃), the expression level of hsp70 gene reached the maximum (10.18 times of the control) in 30 minutes, and then decreased gradually. It reached the minimum (2.85 times of the control) in 180 minutes. Under low temperature stress (4℃), the expression level of hsp70 gene reached the minimum (0.25 times of the control) in 2 hours, and then increased gradually. It keep stable (1.4-1.5 times of the control) in l0 hours. Under UV stress (253.7nm wavelength), the expression level of hsp70 gene reached the maximum (2.33 times of the control) in 3 minutes, and then decreased rapidly. It keep stable (0.2 times of the control) in 60 minutes. It was inferred that hsp70 gene in Beauveria bassiana might play an important role in this three stress tolerances. The results also showed that hsp70 gene promoter of Beauveria bassiana could make gone high express in stress conditions. Therefore, it may have great application value in construction of engineered strain with enhanced anti-adversity ability.