文档介绍:
砂梨离体植物热激蛋白基因的全长克隆及
序列分析#
王俊珍,王利平,洪霓,王**
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(华中农业大学植物科学技术学院)
摘要:热处理是脱除植物病毒常用的技术措施,为明确热处理对植物基因表达的影响及其与
病毒脱除的关系,本研究室前期以砂梨“黄花”离体植株为材料,采用抑制性差减杂交
(Suppression subtractive hybridization, SSH)技术,构建了热处理诱导的梨离体植株基因表
达的抑制性差减文库,筛选出 63 个上调表达和 86 个下调表达的 ESTs,其中热激蛋白(heat
shock protein 70, HSP70)基因表现出明显的上调表达。本研究采用重叠延伸的方法从砂梨“黄
花”离体植株中获得了 HSP70 全长序列为 1950 bp,编码 650 个氨基酸。序列测定及分析表
明,该基因与来源于中国的苹果的热激蛋白分离物同源性高达 %,该基因序列包含了真
核生物 HSP70 基因高度保守的 3 个家族标签,属于 HSP70 家族成员。本研究首次从砂梨克
隆 HSP70 基因,该研究结果为进一步研究热激蛋白基因功能提供材料,也为研究热处理脱
除梨病毒机理提供信息。
关键词:热处理;砂梨;热激蛋白
中图分类号:
Sequence analysis of Heat shock protein 70 gene cloned from
in vitro sand pear plant by thermotherapy
Wang Junzhen, Wang Liping, Hong Ni, Wang Guoping
(College of Plant Science and Technology, Huazhong Agricultural University)
Abstract: Thermotherapy used widely a technique for elimination virus from plants. To determine
the effect of thermotherapy on the relationship of gene expression quantity and virus titers, two
suppression subtractive libraries were constructed from in vitro pear (pyrus pyrifolia)“huanghua”
shoots under 37℃. 149 ESTs were screened including of 63 up-regulated expression genes and 86
down-regulated expression genes. Heat shock protein 70 (HSP70) showed up-regulated expression
among 63 genes. The sequences from HSP70 genes in Pyrus pyrifolia “huanghua” plant shoots
wer