家蚕小热激蛋白家族成员Hsp23.7基因的克隆与分析
Cloning and analysis of small heat-shock gene Hsp23.7 from Bombyx mori
罗素娟1, 2,杨慧鹏1, 2 ,李轶女2,张志芳2,张耀洲1***
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作者单位:1.浙江理工大学生物化学研究所 杭州 310018;2.中国农业科学院生物技术研究所 北京 100081
中文关键词:家蚕,Hsp23.7基因,小热休克蛋白家族,组织表达谱,实时定量PCR
英文关键词: heat shock protein family,tissue expression pattern, Real-time PCR Bombyx mori,Bm-Hsp23.7 gene,small
中文摘要: Hsp23.7基因是小热激蛋白家族的成员,本文研究了家蚕Bombyx mori L. 的Hsp23.7基因,并对其进行了原核表达,获得了相应分子量的表达产物。推导的开放阅读框编码210个氨基酸,分子量为23.7 ku,等电点为5.17。同时,利用实时定量PCR技术对Hsp23.7基因在家蚕不同组织的表达谱进行了鉴定。结果显示Hsp23.7基因在5龄幼虫时期的各组织中都有表达,在卵巢中表达量最高,达到3.64×107拷贝数/μg,其次在脂肪体,翅原基,马氏管中表达量也较高,在血淋巴中表达量最低,仅为7.11×103拷贝数/μg。
英文摘要: Hsp23.7 is one of the small heat-shock family of proteins. The Hsp23.7 gene of Bombyx mori L. (Bm-Hsp23.7) was cloned and expressed in E. coli. The candidate ORF of Bm-Hsp23.7 contains 210 amino acids. Its molecular weight (mw) is 23.7 ku and isoelectric point (pI) is 5.17. Real-time PCR data was used to reveal the profile of the spatio-temporal expression pattern of Bm-Hsp23.7. Absolute quantization demonstrated that Bm-Hsp23.7 was expressed at different levels in all tissues of 5th instar larva of B. mori. The highest level of expression was in the ovary (3.64×107 copy numbers/μg), followed by the fat bodies, then the wing discs and malpighian tubules. The lowest level of expression was in hemocytes (7.11×103 copy numbers/μg).