【摘要】 采用以β-actin基因为内参基因的实时荧光定量PCR方法,检测了不同大小近江牡蛎(Crassostrea hongkongensis)被100μg/L Cu2+处理48 h后闭壳肌、鳃、外套膜、消化腺等四种组织器官中HSC70基因mRNA的表达水平。结果显示,大个体的外套膜、鳃和消化腺中HSC70基因mRNA表达量均明显高于小个体,而在闭壳肌中大小个体表达量无显著差异。同时用不同浓度的Cu2+(0.01、0.1、1,10、50、100μg/L)处理个体大小相似的近江牡蛎不同时间(6 h,2 d,4 d,8 d,20 d,35 d和45 d)后,检测牡蛎外套膜、鳃和消化腺中HSC70基因的表达水平,结果显示用Cu2+连续浸泡处理时,近江牡蛎外套膜和鳃中可诱导HSC70基因显著高表达的Cu2+临界浓度在1~10μg/L范围内,消化腺中可诱导HSC70基因显著高表达的Cu2+临界浓度在0.1~1μg/L范围内。在可诱导HSC70基因表达的浓度组中,在鳃和消化腺中显著高表达峰值出现在第2 d,到第8 d恢复至对照水平;而外套膜中表达峰值出现在第4 d,但延续时间最长可至第20 d。由此可见,近江牡蛎HSC70基因对Cu2+的刺激反应非常灵敏,且持续时间较长,可作为Cu2+污染的早期预警分子。更多还原

【Abstract】 The transcription changes of HSC70 gene of the oyster Crassostrea hongkongensis was quantified by means of real-time fluorescence quantitative RT-PCR method.The housekeeping gene β-actin was used as control gene.The different sizes of oysters which divided into two groups,one was 5~6 cm in length,and the other was 10~12 cm in length,were treated in 100 μg/L Cu2+ solution for 48 hours,and then the transcription changes of HSC70 gene was quantified in the gill,mantle,digestive gland and adductor muscle.The results showed that the expression levels in larger individuals of the oysters were significantly higher than smaller ones in mantle,gill and digestive gland,but not significant difference in the adductor muscle.When the similar size oysters were exposed to 0.01,0.1,1,10,50 and 100 μg/L Cu2+ solution for 6 h,2 d,4 d,8 d,20 d,35 d and 45 d,respectively,the threshold induction concentration of Cu2+ for HSC70 mRNA expression was in the range of 1~10 μg /L in the gill and mantle,and 0.1~1 μg /L in the digestive gland.The oyster HSC70 mRNA in the groups could induce,the HSC70 mRNA levels were significantly increased and reached the peak in gills and digestive gland for 2 days,and peaked for 4 days in mantle after Cu2+ exposure,and then dropped to control levels for 8 days or 20 days.In conclusion,the HSC70 gene response of C.hongkongensis,was sensitive to Cu2+ and could last for a longer time.Therefore,it could be used as a biomarker in response to Cu2+ pollution.更多还原