IDENTIFICATION AND CHARACTERIZATION OF CDNA CLONES ENCODING PLANT CALRETICULIN IN BARLEY
IDENTIFICATION AND CHARACTERIZATION OF CDNA CLONES ENCODING PLANT CALRETICULIN IN BARLEY
复制标题
DOI：
10.1105/tpc.6.6.835
复制
发表时间：
1994-06-01
期刊：
影响因子：
11.6
通讯作者：
PAN, AH
中科院分区：
文献类型：
--
作者：
CHEN, FQ;HAYES, PM;PAN, AH
通过差异筛选，从大麦子房cDNA文库中分离到两个与动物内质网中主要的钙储存蛋白calreticulin同源的cDNA克隆CRH 1和CRH 2，这两个克隆在3'端非翻译区和5'端编码一个推测的N端信号序列的区域存在差异。CRH 1定位于1号染色体负臂。CRH2定位于2号染色体的负臂。推导的氨基酸序列与动物钙网蛋白有50%~ 55%的同源性，并具有相同的三带特征。重组蛋白质用双极-elf染成蓝色，当转移到硝酸纤维素膜上时结合Ca-45（2+）。在卵巢提取物中鉴定出一种分子量约为55 kD的天然蛋白。授粉后1天和胚胎发生早期，在子房中观察到基因表达升高。CRH 1的表达水平高于CRH2。这些研究证实了钙网蛋白在植物细胞中的存在及其在受精过程中的发育调节作用。
Two cDNA clones (CRH1 and CRH2) homologous to animal calreticulin, a major calcium storage protein in the lumen of the endoplasmic reticulum, were isolated from an ovary cDNA library of barley through differential screening, the two clones differ in the 3' untranslated reg ion and the 5' region that encodes a putative N-terminal signal sequence. CRH1 was mapped to the minus arm of chromosome 1. CRH2 was mapped to the minus arm of chromosome 2. The deduced amino acid sequences share 50 to 55% identity with animal calreticulins and exhibit the same three-zone characteristic. Recombinant protein stained blue with Stains-elf and bound Ca-45(2+) when transferred to nitrocellulose membranes. A native protein of similar to 55 kD was identified in ovary extract. Elevated gene expression was observed in ovaries 1 day after pollination and during early embryogenesis. CRH1 was expressed at a higher level than CRH2. These studies demonstrate the presence of calreticulin in plant cells and its developmental regulation in fertilization.