Küster H, Frühling M, Pühler A, Perlick A M
Lehrstuhl für Genetik, Universität Bielefeld, Germany.
Mol Gen Genet. 1996 Oct 28;252(6):648-57. doi: 10.1007/BF02173970.
The broad bean late nodulins, Nvf-28/32, are composed of two types of repetitively occurring sequence modules flanked by unique N- and C-terminal modules. Six isoforms of these nodulins were characterized by a specific modular structure resulting from a different individual order of repetitive sequence modules. A detailed analysis of genomic PCR fragments revealed that the repetitive modules and the N-terminal unique module exactly corresponded to exons, whereas the C-terminal module was specified by two exons. Since those exons encoding the repetitive modules missing in specific Nvf-28/32 isoforms were consistently present within genomic sequences, a post-transcriptional generation of VfNOD28/32 transcripts specifying six Nvf-28/32 nodulins was concluded. Using tissue-print hybridizations, these transcripts were localized in the interzone II-III and the nitrogen-fixing zone III of root nodules. From this and from cDNA-cDNA hybridizations demonstrating a comparable timing of expression of VfNOD28/32 and of leghemoglobin transcripts in root nodules, a function of the modular nodulins Nvf-28/32 in late developmental stages of broad bean nodules was inferred.
蚕豆晚期根瘤蛋白Nvf - 28/32由两种重复出现的序列模块组成,两侧是独特的N端和C端模块。这些根瘤蛋白的六种同工型具有特定的模块化结构,这是由重复序列模块的不同个体顺序导致的。对基因组PCR片段的详细分析表明,重复模块和N端独特模块与外显子完全对应,而C端模块由两个外显子指定。由于编码特定Nvf - 28/32同工型中缺失的重复模块的那些外显子始终存在于基因组序列中,因此得出结论,VfNOD28/32转录本的转录后生成指定了六种Nvf - 28/32根瘤蛋白。使用组织印记杂交技术,这些转录本定位在根瘤的II - III区和固氮区III。由此以及通过cDNA - cDNA杂交证明VfNOD28/32和豆血红蛋白转录本在根瘤中的表达时间相当,推断出模块化根瘤蛋白Nvf - 28/32在蚕豆根瘤发育后期的功能。