Sullivan T D, Strelow L I, Illingworth C A, Phillips R L, Nelson O E
Laboratory of Genetics, University of Wisconsin, Madison 53706.
Plant Cell. 1991 Dec;3(12):1337-48. doi: 10.1105/tpc.3.12.1337.
A mutant allele of the maize brittle-1 (bt1) locus, brittle-1-mutable (bt1-m), was shown genetically and molecularly to result from the insertion of a defective Suppressor-mutator (dSpm) transposable element. An Spm-hybridizing restriction enzyme fragment, which cosegregates with the bt1-m allele and is absent from wild-type revertants of bt1-m, was identified and cloned. Non-Spm portions of it were used as probes to identify wild-type (Bt1) cDNAs in an endosperm library. The 4.3-kb bt1-m genomic clone contains a 3.3-kb dSpm, which is inserted in an exon and is composed of Spm termini flanking non-Spm sequences. RNA gel blot analyses, using a cloned Bt1 cDNA probe, indicated that Bt1 mRNA is present in the endosperm of developing kernels and is absent from embryo or leaf tissues. Several transcripts are produced by bt1-m. The deduced translation product from a 1.7-kb Bt1 cDNA clone has an apparent plastid transit peptide at its amino terminus and sequence similarity to several mitochondrial inner-envelope translocator proteins, suggesting a possible role in amyloplast membrane transport.
玉米脆-1(bt1)位点的一个突变等位基因,即脆-1-可变(bt1-m),经遗传学和分子学研究表明是由一个有缺陷的抑制子-突变体(dSpm)转座元件插入所致。鉴定并克隆了一个与bt1-m等位基因共分离且在bt1-m的野生型回复体中不存在的Spm杂交限制性酶切片段。其非Spm部分被用作探针,在胚乳文库中鉴定野生型(Bt1)cDNA。4.3kb的bt1-m基因组克隆包含一个3.3kb的dSpm,它插入到一个外显子中,由侧翼为非Spm序列的Spm末端组成。使用克隆的Bt1 cDNA探针进行的RNA凝胶印迹分析表明,Bt1 mRNA存在于发育中籽粒的胚乳中,而在胚或叶组织中不存在。bt1-m产生了几种转录本。从一个1.7kb的Bt1 cDNA克隆推导的翻译产物在其氨基末端有一个明显的质体转运肽,并且与几种线粒体内膜转运蛋白有序列相似性,这表明它可能在造粉体膜转运中起作用。