Prathepha Preecha
Department of Biotechnology, Faculty of Technology, Mahasarakham University, Mahasarakham Province 44000, Thailand.
Pak J Biol Sci. 2007 Aug 1;10(15):2500-4. doi: 10.3923/pjbs.2007.2500.2504.
The identification of a number of cDNA sequences at the splice sites of Waxy (Wx) gene in rice endosperm point to the variation of gene expression associated with the G to T mutation at the 5' splice site of intron 1 and the alternative splicing produces variants transcripts by shortening. As consequence, variation of amylose content in this tissue. To determine whether the splicing type of Wx genes are also found in rice endosperm of Thai rice cultivars, cDNA of the gene have been isolated by using RT-PCR and sequenced. Analysis of the twenty-five cDNA clones led to the identification of the splicing pattern of the gene, four cryptic splice sites was found in cDNA sequences of low-amylose cultivars (<20%), while two cryptic splice sites of intermediate (20-25%) and high (>25%) amylose cultivars were characterized. The alternatively spliced transcripts were observed in much higher proportion in low amylose cultivars than that in intermediate and high amylose cultivars. These findings raise a possibility explanation that the degree of amylose content in non-glutinous rice cultivars may reflect to the amount of efficiently spliced transcripts.
在水稻胚乳中蜡质(Wx)基因剪接位点处多个cDNA序列的鉴定表明,基因表达的变异与内含子1的5'剪接位点处的G到T突变相关,并且可变剪接通过缩短产生变异转录本。结果,该组织中直链淀粉含量发生变化。为了确定泰国水稻品种的水稻胚乳中是否也存在Wx基因的剪接类型,已通过逆转录聚合酶链反应(RT-PCR)分离该基因的cDNA并进行测序。对25个cDNA克隆的分析导致了该基因剪接模式的鉴定,在低直链淀粉品种(<20%)的cDNA序列中发现了四个隐蔽剪接位点,而中间(20-25%)和高(>25%)直链淀粉品种的两个隐蔽剪接位点得到了表征。在低直链淀粉品种中观察到可变剪接转录本的比例远高于中间和高直链淀粉品种。这些发现提出了一种可能性解释,即非糯性水稻品种中直链淀粉含量的程度可能反映了有效剪接转录本的数量。