新疆梨S28-RNase基因的全长cDNA克隆及序列特征分析

刘敏; 张琳; 王明媚; 谭晓风; 宋志波; 李秀根; 曹玉芬

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Molecular cloning and sequence characterization of the full-length cDNA encoding S28-RNase from Xinjiang pear (Pyrus sinkiangenis)

首发时间：2012-11-07

Liu Min ¹
Liu Min, (1987-), male, Cultivation and breeding of Non-wood Forest Crops.
Zhang Lin ²
Zhang Lin (1978-), male, Ph.D., Associate Professor, Cultivation and breeding of Non-wood Forest Crops
Wang Mingmei ³ Tan Xiaofeng ² Song Zhibo ² Li Xiugen ⁴ Cao Yufen ⁵

1、Key Laboratory of Cultivation and Protection for Non-Wood Forest Trees, Ministry of Education, Central South University of Forestry and Technology, ChangSha 410004
2、Key Laboratory of Cultivation and Protection for Non-Wood Forest Trees, Ministry of Education, Central South University of Forestry and Technology, Changsha 410004
3、Colledge of Forestry, Central South University of Forestry and Technology, Changsha 410004
4、Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Zhengzhou 450009, China
5、Research Institute of Pomology, Chinese Academy of Agricultural Sciences, Xingcheng 125100, China

Abstract：The full length cDNA encoding S28-RNase (GenBank accession No. EF566872) was cloned from styles of Xinjiang pear (Pyrus sinkiangeni) cultivar 'Kuerlexiangli' by Reverse transcription-PCR (RT-PCR) and Rapid Amplification of cDNA Ends (RACE) technology. The S28-RNase cDNA contained 865 nucleotides including a complete open reading frame (ORF) predicted to encode a protein of 228 amino acids. The S28-RNase displayed the basic structural features of pear S-RNases, i.e. five conserved regions (C1, C2, C3, RC4 and C5) and a hypervariable (HV) region. At the deduce amino acid level, S28-RNase showed 58.4% to 99.6% similarity with other pear S-RNases. The isoeletric point (pI) and molecular weight (Mw) of S28-RNase were predicted to be 9.30 and 25.6 kDa, respectively. The S28-RNase showed the conserved motifs with two histidine residue, His-61 and His-117 which are essential for T2/S type RNase activity. It also presented eight cysteine residues mostly conserved in S-RNases, forming four disulfide bridges important for the formation or stabilization of their tertiary structure. The S28-RNase also showed seven potential N-glycosilation sites, of which only one, Asn-145, conserved in rosaceous S-RNases, whose glycans may be important in the folding and stabilization of the core structure.

keywords： Gametophytic self-incompatibility (GSI) 'Kuerlexiangli' Rapid Amplification of cDNA Ends S28-RNase gene

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新疆梨S28-RNase基因的全长cDNA克隆及序列特征分析

刘敏 ¹
Liu Min, (1987-), male, Cultivation and breeding of Non-wood Forest Crops.
张琳 ¹
Zhang Lin (1978-), male, Ph.D., Associate Professor, Cultivation and breeding of Non-wood Forest Crops
王明媚 ² 谭晓风 ¹ 宋志波 ¹ 李秀根 ³ 曹玉芬 ⁴

1、中南林业科技大学经济林培育与保护教育部重点实验室，长沙 410004
2、中南林业科技大学林学院，长沙 410004
3、中国农业科学院郑州果树研究所，河南郑州
4、中国农业科学院果树研究所，辽宁兴城 121600

摘要：通过逆转录PCR及快速扩增cDNA末端技术从新疆梨品种'库尔勒香梨'梨中克隆到S28-RNase基因的全长cDNA序列（GenBank接受号为EF566872）。该cDNA克隆总长865 bp，包含一个完整的开放阅读框，编码228个氨基酸。S28-RNase表现出梨S-RNase基因基本的结构特征，即其具有五个保守区（C1，C2，C3，RC4及C5）和一个高变区。在推导氨基酸水平上，S28-RNase与梨其它S-RNase基因的相似性为58.4％至99.6％。S28-RNase等电点和分子量分别为9.30、25.6 kDa，含有8个高度保守的半胱氨酸残基，形成4个二硫键，含有2个保守的影响S-RNase酶活性的组氨酸残基，RC4区存在一个保守的"Asn-Xaa-Ser/Thr"结构，该结构在核心结构折叠过程中起重要作用。

关键词：配子体自交不亲和库尔勒香梨快速扩增cDNA末端 S28-RNase基因

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Liu Min,Zhang Lin,Wang Mingmei,et al. Molecular cloning and sequence characterization of the full-length cDNA encoding S28-RNase from Xinjiang pear (Pyrus sinkiangenis)[EB/OL]. Beijing:Sciencepaper Online[2012-11-07]. https://www.paper.edu.cn/releasepaper/content/201211-108.

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论文编号	201211-108
论文题目	新疆梨S28-RNase基因的全长cDNA克隆及序列特征分析
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