热休克对小鼠胚胎成纤维细胞CaM mRNA和HSF活性的影响

试验分别以38、40和42℃对原代培养的小鼠胚胎成纤维细胞进行热休克处理,采用半定量PCR方法和凝胶迁移阻滞（EMSA）对CaM mRNA和HSF活性进行检测,并与对照组（37℃）比较,以确定热休克对小鼠胚胎成纤维细胞CaM mRNA和HSF活性的影响。结果表明,38℃热休克处理组小鼠胚胎成纤维细胞的CaMmRNA表达量显著高于对照组（P〈0.05）;40和42℃处理组细胞的CaM mRNA表达量极显著高于37℃组（P〈0.01）,但与40℃处理组相比,42℃处理组细胞的CaM mRNA表达量略有下降。通过凝胶阻滞分析检测HSF与DNA的结合活性,发现与对照组相比,38、40和42℃热休克处理...

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Bibliographic Details
Published in东北农业大学学报 Vol. 42; no. 6; pp. 71 - 74
Main Author 李金波 张霞 田文儒 曹荣峰 高善颂 张桂林
Format Journal Article
LanguageChinese
Published 青岛农业大学动物科技学院,山东青岛,266109 2011
Subjects
Ca2
CaM
HSF
热休克
HSF
Ca2
热休克
CaM
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ISSN1005-9369
DOI10.3969/j.issn.1005-9369.2011.06.014

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Summary:试验分别以38、40和42℃对原代培养的小鼠胚胎成纤维细胞进行热休克处理,采用半定量PCR方法和凝胶迁移阻滞（EMSA）对CaM mRNA和HSF活性进行检测,并与对照组（37℃）比较,以确定热休克对小鼠胚胎成纤维细胞CaM mRNA和HSF活性的影响。结果表明,38℃热休克处理组小鼠胚胎成纤维细胞的CaMmRNA表达量显著高于对照组（P〈0.05）;40和42℃处理组细胞的CaM mRNA表达量极显著高于37℃组（P〈0.01）,但与40℃处理组相比,42℃处理组细胞的CaM mRNA表达量略有下降。通过凝胶阻滞分析检测HSF与DNA的结合活性,发现与对照组相比,38、40和42℃热休克处理明显增加了HSF活性。
Bibliography:heat shock; Ca2＋; CaM; HSF
23-1391/S
LI Jinbo,ZHANG Xia,TIAN Wenru,CAO Rongfeng,GAO Shansong,ZHANG Guilin（College of Animal Science and Veterinary Medicine,Qingdao Agricultural University,Qingdao Shandong 266109,China）
This study was carried out to detecte the expression of CaM mRNA and the activity of HSF by using the methods of RT-PCR and Gel mobility shift assay（EMSA） after the primary cultured mouse embryonic fibroblasts were exposed to 38,40 and 42 ℃.The results showed that the expressions of CaM mRNA in 38℃ groups was significantly higher than that in the control group（P0.05）.However,the expression of CaM mRNA in the group treated at 40 and 42 ℃ were extremely higher compared to that of the control group（P0.01）;The expressions of CaM mRNA in the groups treated at 42 ℃ was lower compared with that in 40 ℃ groups.Compared to the control group,the activity of HSF in the heat shock groups was significantly increased as determined by EMSA.
ISSN:1005-9369
DOI:10.3969/j.issn.1005-9369.2011.06.014