Expression of the β-subunit of β-conglycinin in seeds of transgenic plants-猫眼课题宝

β-伴大豆球蛋白的β-亚基在转基因植物种子中的表达

DOI：

发表时间：

1987

期刊：

Planta

影响因子：

4.3

通讯作者：

中科院分区：

生物学2区

文献类型：

作者：

E. Bray;S. Naito;N. Pan;E. Anderson;P. Dubé;R. Beachy

大豆（Glycine max（L.）梅尔）种子含有由多基因家族编码的贮藏蛋白β-伴大豆球蛋白。β-伴大豆球蛋白由α′、α和β三个亚基组成。β-伴大豆球蛋白的β-亚基的基因组克隆已经通过限制酶图谱和杂交选择体外翻译随后免疫沉淀来表征。为了研究该β亚基基因的发育调控，以转基因矮牵牛和烟草种子为材料，研究了该基因的表达。植物在矮牵牛和烟草种子中表达的β-亚基可被抗β-伴大豆球蛋白血清识别，相对分子质量为53000，与天然蛋白相当。通过等电聚焦、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹分析分离矮牵牛种子蛋白，结果表明产生了多种等电形式的β亚基。在转基因矮牵牛植物的成熟种子中，β-亚基蛋白的积累存在大约两倍的变化，每个转基因矮牵牛植物含有单个β-亚基基因。但成熟种子中蛋白质的积累水平与发育种子中β亚基mRNA的含量不相关。转基因矮牵牛种子中β-亚基蛋白的积累量低于含有单一α′-亚基基因的转基因矮牵牛种子中α′-亚基蛋白的积累量，也低于含有8-13个β-亚基基因的成熟大豆种子中β-亚基蛋白的积累量。在转基因烟草植物中，种子中β-亚基蛋白的积累通常与掺入不同转化体中的基因的数量密切相关。

Soybean (Glycine max (L.) Merr.) seeds contain the storage protein β-conglycinin, encoded by a multigene family. β-Conglycinin consists of three subunits; α′, α, and β. A genomic clone for a β-subunit of β-conglycinin has been characterized by restriction-enzyme mapping and hybrid selected in-vitro translation followed by immunoprecipitation. In order to determine the developmental regulation of this β-subunit gene, its expression was studied in seeds of transgenic petunia (Petunia hybrida) and tobacco (Nicotiana tabacum L.) plants. The β-subunit expressed in seeds of petunia and tobacco was recognized by anti-β-conglycinin serum at a relative molecular mass of 53 000, equivalent to that of the native protein. Separation of the petunia-seed proteins by isoelectric focusing followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis showed that multiple isoelectric forms of the β-subunit were produced. There was approximately a twofold variation in the accumulation of the β-subunit protein in the mature seeds of transgenic petunia plants, each containing a single β-subunit gene. However, the level of protein accumulation in mature seeds and the amount of β-subunit mRNA in developing seeds was not correlated. Accumulation of the β-subunit protein in transgenic seeds was less than the α′-subunit protein that accumulated in transgenic petunia seeds containing a single α′-subunit gene and less than the amount of the β-subunit in mature soybean seeds which contain 8–13 β-subunit genes. In transgenic tobacco plants, the accumulation of the β-subunit protein in seeds was generally well correlated with the number of genes that were incorporated in the different transformants.

Immunoautoradiographic detection of proteins after electrophoretic transfer from gels to diazo-paper: analysis of adenovirus encoded proteins.