小麦硬度主效基因Pina和Pinb的克隆和序列分析

籽粒硬度是小麦品质改良的重要目标性状. 根据已报道的谷类作物中硬度主效基因Pina和Pinb的DNA序列, 设计了两对简并引物, 以我国软质小麦品种京411基因组DNA为模板, 进行PCR扩增, 分别获得了约450 bp的Pina和Pinb基因的全长特异性片段. 将其克隆到pGEM-3Zf(+)上, 重组子和酶切鉴定正确后, 进行序列测定和分析. 结果表明, 与

Cloning and Sequence Analysis of Pina and Pinb Genes Controlling Grain Hardness in Common Wheat

Grain hardness is one of the most important characters in wheat quality improvement. Based on the homologues of Pin a and Pin b genes, which are major genes controlling the grain hardness in cereal crops, two pairs of degenerate primers were designed. The specific fragments of about 450 bp in size were obtained, respectively, after PCR amplification using the genomic DNA of wheat variety Jing411 as template. Then they were cloned into vector pGEM 3Zf( ), and sequenced after the identification of the recombinants and endonuclease analysis. The results indicated that, compared with the Pin a gene from wheat variety Heron, the Pin a gene in Jing411 shared 98.9% and 98% homology in nucleotide acid sequence and amino acid sequence, respectively. The whole length of Pin a gene was 447 bp, encoding 149 amino acid residues, having the signal peptide of 19aa and the tryptophan rich domain (WWKWWK) which are specific to Pin a gene in cereal crops. Similarly, the Pin b gene shared 99.8% and 99.3% homologies in nucleotide acid sequence and amino acid sequence respectively compared with the cloned Pin b gene in soft wheat variety Heron. The whole length of Pin b gene was 447 bp, encoding 149 amino acid residues, having the signal peptide of 19aa and the tryptophan rich domain (KWWK) which were specific to Pin b gene in cereal crops. The isolation of Pin a and Pin b gene laid a fundamental basis for further improvement of the grain hardness of wheat variety through genetic engineering.