USDA/Agricultural Research Service and the Department of Agronomy, Purdue University, 47907, West Lafayette, IN, USA.
Theor Appl Genet. 1990 Jun;79(6):785-92. doi: 10.1007/BF00224246.
Soybean vegetative storage proteins (VSPs) were purified and characterized. Anion exchange HPLC resolved partially purified VSPs into fractions containing 27-kD/27-kD and 29-kD/29-kD homodimers and 27-kD/29-kD heterodimers. Reversed-phase HPLC resolved partially purified VSPs into three fractions. One fraction contained only 27-kD VSP and the other two contained 29-kD VSP. The two 29-kD VSP fractions differed with respect to their cyanogen bromide cleavage patterns, an observation that indicated the 29-kD VSPs were heterogeneous. Genomic clones that contained 29-kD VSP genes were also isolated and characterized. One genomic clone contained a complete 29-kD VSP gene and was sequenced. The coding region in the clone contained two introns whose borders had regulatory sequences typical of other eukaryotic genes. Putative polyadenlyation signals were present in the 3'-flanking region of the gene, while putative TATA, CAAT, and enhancer core sequences were found in the 5'-flanking regions. A second genomic clone that was studied contained the 5' regions of two partial 29-kD VSP genes in an inverted linkage. Genomic DNA gel blots showed that the two genes were organized in the same arrangement in the soybean genome.
大豆营养体贮藏蛋白（VSP）被分离和鉴定。阴离子交换 HPLC 将部分纯化的 VSP 分离成含有 27kD/27kD 和 29kD/29kD 同二聚体和 27kD/29kD 异二聚体的组分。反相 HPLC 将部分纯化的 VSP 分离成三个组分。一个组分仅含有 27kD VSP，其它两个组分含有 29kD VSP。两个 29kD VSP 组分的氰化溴裂解图谱不同，这表明 29kD VSP 是不均一的。还分离和鉴定了含有 29kD VSP 基因的基因组克隆。一个基因组克隆含有完整的 29kD VSP 基因并被测序。该克隆中的编码区含有两个内含子，其边界具有其它真核基因典型的调节序列。在基因的 3′侧翼区存在潜在的多聚腺苷酸化信号，而在 5′侧翼区发现了潜在的 TATA、CAAT 和增强子核心序列。研究的第二个基因组克隆含有两个部分 29kD VSP 基因的 5′区域，以反向连接的方式排列。基因组 DNA 凝胶印迹显示，这两个基因在大豆基因组中以相同的方式排列。
