水稻杂种与亲本间差异表达cDNA片段S600的分离克隆
Identification and Cloning of S600 cDNA Fragment That Is Differentially Expressed in a Rice Hybrid and Its Parents
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摘要: 采用cDNA-AFLP技术分离克隆了水稻杂种与亲本间差异表达基因片段S600。Northern杂交结果表明:在分蘖期和始穗期,S600在杂种和父本中表达丰度均较高,而在母本中表达丰度相对较低。S600在分蘖期和始穗期表达量不同,暗示了该基因的表达还受到发育时期的调节。同源搜索结果表明S600片段是水稻SBPase的部分编码序列。为了获得完整编码序列,以S600序列检索粳稻日本晴cDNA数据库,获得了两个高度同源(99%)且功能未知的全长cDNA克隆(AK062089和AK065773)。序列分析表明它们均包含一个相同的1179bp的开放阅读框,编码392个氨基酸组成的水稻SBPase前体,其中包含有与底物结合、氧化还原调节有关的保守氨基酸残基。检索发现该基因在水稻日本晴基因组中只有单个座位。Abstract: A cDNA fragment S600 that shows differential expression pattern between a rice hybrid and its parental lines was identified by cDNA-AFLP. Northern blot results showed that the transcript level of S600 in female parent is lower than that in hybrid or male parent at both tillering and flowering stages. The transcript level in the two stages is different, indicating this gene is developmental-regulated. Homology search revealed that S600 is a part of coding sequence of rice sedoheptulose-J, 7-bisphosphatase. To obtain its complete coding sequence, S600 sequence was used to BLAST against Nipponbare cDNA database. Two highly homologous (99% identities) and functionally unknown cDNA clones (AK062089 and AK065773) were identified. Sequence analysis showed that both of the clones contain an identical open reading frame of (1179) bp that encodes 392 AAs SBPase precursor. The conservative amino acid residues involved in substrate binding and redox-regulation were found in the peptide. BLAST search result also indicates that SBPase gene is single locus in Nipponbare genome.