Podkowinski J, Sroga G E, Haselkorn R, Gornicki P
Department of Molecular Genetics and Cell Biology, University of Chicago, IL 60637, USA.
Proc Natl Acad Sci U S A. 1996 Mar 5;93(5):1870-4. doi: 10.1073/pnas.93.5.1870.
An entire gene encoding wheat (var. Hard Red Winter Tam 107) acetyl-CoA carboxylase [ACCase; acetyl-CoA:carbon-dioxide ligase (ADP-forming), EC 6.4.1.2] has been cloned and sequenced. Comparison of the 12-kb genomic sequence with the 7.4-kb cDNA sequence reported previously revealed 29 introns. Within the coding region, the exon sequence is 98% identical to the known wheat cDNA sequence. A second ACCase gene was identified by sequencing fragments of genomic clones that include the first two exons and the first intron. Additional transcripts were detected by 5' and 3' RACE analysis (rapid amplification of cDNA ends). One set of transcripts had a 5' end sequence identical to the cDNA found previously and another set was identical to the gene reported here. The 3' RACE clones fall into four distinguishable sequence sets, bringing the number of ACCase sequences to six. None of these cDNA or genomic clones encodes a chloroplast targeting signal. Identification of six different sequences suggests that either the cytosolic ACCase genes are duplicated in the three chromosome sets in hexaploid wheat or that each of the six alleles of the cytosolic ACCase gene has a readily distinguishable DNA sequence.
编码小麦(品种:硬红冬小麦Tam 107)乙酰辅酶A羧化酶[ACCase;乙酰辅酶A:二氧化碳连接酶(ADP形成),EC 6.4.1.2]的完整基因已被克隆并测序。将12 kb的基因组序列与先前报道的7.4 kb cDNA序列进行比较,发现有29个内含子。在编码区内,外显子序列与已知的小麦cDNA序列有98%的同一性。通过对包含前两个外显子和第一个内含子的基因组克隆片段进行测序,鉴定出了第二个ACCase基因。通过5'和3' RACE分析(cDNA末端快速扩增)检测到了其他转录本。一组转录本的5'末端序列与先前发现的cDNA相同,另一组与本文报道的基因相同。3' RACE克隆分为四个可区分的序列组,使ACCase序列的数量达到六个。这些cDNA或基因组克隆均未编码叶绿体靶向信号。六个不同序列的鉴定表明,要么细胞质ACCase基因在六倍体小麦的三个染色体组中发生了重复,要么细胞质ACCase基因的六个等位基因各自具有易于区分的DNA序列。