Nuccio M L, Thomas T L
Department of Biology, Texas A&M University, College Station, 77843-3258, USA.
Plant Mol Biol. 1999 Apr;39(6):1153-63. doi: 10.1023/a:1006101404867.
A modified protocol for differential display of mRNA was used to identify and clone genes expressed in developing Arabidopsis thaliana seeds. Two novel embryo-specific genes designated ATS1 and ATS3 (Arabidopsis thaliana seed gene) were identified. In situ hybridization showed that, spatially, ATS1 is expressed in a pattern similar to the Arabidopsis GEA1 gene and that ATS3 is expressed in a pattern similar to the Arabidopsis seed storage protein genes. Southern analysis of Arabidopsis genomic DNA indicated that ATS1 is a member of a small gene family and that ATS3 is present as a single copy in the diploid genome. Sequence analysis of both genes showed that ATS1 is similar to the rice EFA27 gene and that ATS3 is unique. Western analysis and light level immunocytochemistry using antisera raised against the putative ATS1 and ATS3 translation products verified that ATS1 and ATS3 proteins are seed-specific and accumulate in a spatial pattern similar to their respective transcripts. Taken together, these data show that ATS1 and ATS3 are novel embryo-specific genes in Arabidopsis.
采用一种改良的mRNA差异显示方案来鉴定和克隆在拟南芥种子发育过程中表达的基因。鉴定出了两个新的胚胎特异性基因，命名为ATS1和ATS3（拟南芥种子基因）。原位杂交显示，在空间上，ATS1的表达模式与拟南芥GEA1基因相似，而ATS3的表达模式与拟南芥种子贮藏蛋白基因相似。对拟南芥基因组DNA的Southern分析表明，ATS1是一个小基因家族的成员，而ATS3在二倍体基因组中以单拷贝形式存在。对这两个基因的序列分析表明，ATS1与水稻EFA27基因相似，而ATS3是独特的。使用针对推定的ATS1和ATS3翻译产物产生的抗血清进行的Western分析和光镜水平免疫细胞化学证实，ATS1和ATS3蛋白是种子特异性的，并且以与其各自转录本相似的空间模式积累。综上所述，这些数据表明ATS1和ATS3是拟南芥中新型的胚胎特异性基因。
