| 摘要: |
| 【目的】从鸭梨中克隆GAPDH基因家族，筛选合适的鸭梨内参基因。【方法】采用同源克隆的方法，克隆鸭梨GAPDH基因家族，用半定量PCR方法分析4个鸭梨GAPDH基因在鸭梨幼叶、花蕾、盛花期花瓣、幼果期果肉和成熟期果心，以及幼果期、膨大期、成熟期和褐变期果心的表达特征。【结果】从鸭梨中成功克隆出了4个GAPDH基因，分别命名为PbGAPDHa、PbGAPDHb、PbGAPDHc1和PbGAPDHc2。其中PbGAPDHa和PbGAPDHb是由一个共同的祖先基因倍增进化来的，主要在鸭梨的幼叶中表达，其蛋白定位在质体；而PbGAPDHc1和PbGAPDHc2是一个基因的2种拷贝形式，在鸭梨的花、幼叶、果实等组织中表达量基本一致，在不同发育时期的果心中表达量也相近，其蛋白定位在细胞质。【结论】PbGAPDHc1和PbGAPDHc2适合作为鸭梨的内参基因。 |
| 关键词: 鸭梨 3-磷酸甘油醛脱氢酶 同源基因克隆 半定量分析 基因进化 |
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| 基金项目:现代农业（梨）产业技术体系建设专项(CARS-29) |
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| Cloning and characters analysis of GAPDH gene family in Yali pear |
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| Abstract: |
| 【Objective】This study was to clone GAPDH genes family and screen ideal control gene in ‘Yali’ pear.【Method】Homologous cloning method was usd to clone GAPDH gene family and the expression patterns of four genes in young leaves,buds,petals,flesh,core and four main development stage core of young fruit,expension fruit,mature fruit and browning fruit were analyzed through semi-quantility PCR.【Result】In this study,four GAPDH gene family members were isolated from ‘Yali’ pear and named as PbGAPDHa,PbGAPDHb,PbGAPDHc1 and PbGAPDHc2.PbGAPDHa and PbGAPDHb were evolved from the duplication of a old ancstor,mainly expressed in pear leaves,and two putative proteins were located in plastid.PbGAPDHc1 and PbGAPDHc2 became two copies from one old GAPDHc after dicotyledonous seperation and two putative proteins were located in cytoplasm.They were expressed constitutively in all tested organs,including leaf,bud,pestal,fruit flesh,core fuit.There was no significant difference in mRNA expression of PbGAPDHc1 and PbGAPDHc2 in different core fruit development stages.【Conclusion】PbGAPDHc1 and PbGAPDHc2 can be used as control gene for other gene expression study in ‘Yali’ pear. |
| Key words: ‘Yali’ pear glyceraldehyde-3-phosphate dehyrogenase(GAPDH) homologous cloning semi-quantity analysis gene evolution |
