Single nucleotide polymorphism genotyping of the barley waxy gene by polymerase chain reaction with confronting two-pair primers
Single nucleotide polymorphism genotyping of the barley waxy gene by polymerase chain reaction with confronting two-pair primers
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DOI：
10.1111/j.1439-0523.2004.00970.x
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发表时间：
2004-06-01
期刊：
影响因子：
2
通讯作者：
Takeda, K
中科院分区：
文献类型：
--
作者：
Domon, E;Yanagisawa, T;Takeda, K
采用高通量单核苷酸多态性（SNP）基因分型方法筛选无直链淀粉的大麦突变体，该突变体的蜡质基因在第5外显子中具有C- to -t碱基替换，将野生型基因的Gln-89转化为终止密码子。对携带无直链淀粉蜡质基因的F-2群体进行分离检查。聚合酶链反应与对抗性双对引物（PCR- ctpp）产生的等位基因特异性PCR产物具有不同的大小，并以共显性方式遗传。利用pcr - cttp方法，在亲本株中正确鉴定出两个具有SNP的大麦蜡质基因等位基因。PCR-CTPP在F-2群体中检测到的SNP分离符合预期的1:2:1比例。在标记辅助选择中，PCR-CTPP程序可以为衍生的裂解扩增多态性序列提供一种节省时间和成本效益的替代方法。
A high-throughput single nucleotide polymorphism (SNP) genotyping procedure was developed to select amylose-free barley mutants whose waxy genes had a C- to T-base substitution in exon 5, which converted Gln-89 of the wild-type gene into a termination codon. An F-2 population carrying an amylose-free waxy gene was checked for segregation. Polymerase chain reaction with confronting two-pair primers (PCR-CTPP) produced allele-specific PCR products that have different sizes and are inherited in a co-dominant manner. Two alleles of the barley waxy gene with SNP were correctly identified in parental strains using the PCR-CTPP procedure. Segregation of the SNP as detected by PCR-CTPP in an F-2 population fitted the expected 1 : 2 : 1 ratio. The PCR-CTPP procedure can provide a time saving and cost-effective alternative to derived cleaved amplified polymorphic sequence in marker-assisted selection.