Construction of Stress Induced Full Length cDNA Library of Sedum alfredii and Isolation of Genes Related to Cd-tolerance

LIU Ming-ying; QIAO Gui-rong; JIANG Jing; QIU Wen-min; ZHUO Ren-ying

A cadmium stress induced full-length cDNA Library from the Sedum alfredii was constructed by using improved SMART cDNA library construction protocol. Total RNA were extracted from S. alfredii under cadmium stress. After synthesizing the first-strand cDNA using limited amount of total RNA, the double-strand cDNA was amplified with long-distance PCR method. The full-length cDNA fragment was linked to versatile vector pYES2.0G, then the linked product was transferred into yeast INVSc1. The clones were randomly selected to sequence and bioinformatics analysis was carried out. The inserted fragments in the library are about 1000 bp length. This full-length cDNA Library provides a important tool for the study of the salt tolerance mechanisms of S. alfredii.

1. Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, Zhejiang, China

2. State Key Laboratory of Tree Genetics and Breeding, Beijing 100091, China

Received Date: 2011-10-11

Available Online: 2012-06-13

Abstract: A cadmium stress induced full-length cDNA Library from the Sedum alfredii was constructed by using improved SMART cDNA library construction protocol. Total RNA were extracted from S. alfredii under cadmium stress. After synthesizing the first-strand cDNA using limited amount of total RNA, the double-strand cDNA was amplified with long-distance PCR method. The full-length cDNA fragment was linked to versatile vector pYES2.0G, then the linked product was transferred into yeast INVSc1. The clones were randomly selected to sequence and bioinformatics analysis was carried out. The inserted fragments in the library are about 1000 bp length. This full-length cDNA Library provides a important tool for the study of the salt tolerance mechanisms of S. alfredii.

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[1]	[[14] Du J, Huang Y P, Xi J, et al. Functional gene-mining for salt-tolerance genes with the power of Arabidopsis [J]. Plant J, 2008, 56(4): 653-664
[2]	马瑾, 潘根兴, 万洪富, 等. 珠江三角洲典型区域土壤重金属污染探查研究[J]. 土壤通报, 2004, 35(5): 636-638
[3]	杨肖娥, 龙新宪, 倪吾钟, 等. 东南景天(Sedum alfredii H) ——一种新的锌超积累植物[J]. 科学通报, 2002, 47(13): 1003-1006
[4]	熊愈辉, 杨肖娥, 叶正钱, 等. 东南景天对镉、铅的生长反应与积累特性比较[J]. 西北农林科技大学报: 自然科学版, 2004, 32(6): 101-106
[5]	熊愈辉. 东南景天对镉的耐性生理机制及其对土壤镉的提取与修复作用的研究[D]. 杭州:浙江大学,2005
[6]	龙新宪, 王艳红, 刘洪彦. 不同生态型东南景天对土壤中Cd的生长反应及吸收积累的差异性[J]. 植物生态学报, 2008, 32(1): 168-175
[7]	Ichikawa T, Nakazawa M, Kawashima M, et al. The FOX hunting system: an alternative gain-of-function gene hunting technique [J]. Plant J, 2006, 48(6):974-985
[8]	Frommer W B, Hummel S, Riesmeier J W. Expression cloning in yeast of a cDNA encoding a broad specificity amino acid permease from Arabidopsis thaliana [J]. Proc Natl Acad Sci USA, 1993, 90(13): 5944-5948
[9]	Yamada A, Saitoh T, Mimura T, et al. Expression of mangrove allene oxide cyclase enhances salt tolerance in Escherichia coli, yeast and tobacco cells [J]. Plant ＆ Cell Physiol, 2002, 43(8):903-910
[10]	Yamada A,Tsutsumi K,Tanimoto S, et al. Plant RelA/SpoT homolog confers salt tolerance in Escherichia coli and Saccharomyces cerevisiae [J]. Plant and Cell Physiology, 2003, 44(1): 3-9[ZK)]
[11]	Zhang Z L, Zhang S P, Zhang Y, et al. Arabidopsis floral initiator SKB1 confers high salt tolerance by regulating transcription and pre-mRNA splicing through altering histone H4R3 and small nuclear ribonucleoprotein LSM4 methylation [J]. The Plant Cell, 2011, 23(1): 396-411
[12]	Wu Y, Zhao Q, Gao L, et al. Isolation and characterization of low-sulphur-tolerant mutants of Arabidopsis [J]. Journal of Experimental Botany, 2010, 61(12): 3407-3422
[13]	Monteiro C C, Carvalho R F, Grato P L, et al. Biochemical responses of the ethyleneinsensitive Never ripe tomato mutant subjected to cadmium and sodium stresses [J]. Environmental and Experimental Botany, 2011, 71(2): 306-320
[14]	Lee J H, Montagu M V, Verbruggen N. A highly conserved kinase is an essential component for stress tolerance in yeast and plant cells [J]. Proc Natl Acad Sci USA, 1999, 96(10): 5873-5877
[15]	Soares M B, Bonaldo M F, Jelene P, et al. Construction and characterization of a normalized cDNA library [J]. Proc Natl Acad Sci USA 1994, 91(20): 9228-9232
[16]	LeClere S, Bartel B. A library of Arabidopsis 35S-cDNA lines for identifying novel mutants [J]. Plant Mol Biol, 2001, 46(6):695-703
[17]	Fischer K, Holt D C, Wilson P, et al. Normalization of a cDNA library cloned in lambda ZAP by a long PCR and cDNA reassociation procedure [J]. Biotechniques, 2003, 34(2):250-252, 254
[18]	Ni W S, Lei Z Y, Chen X, et al. Construction of a plant transformation-ready expression cDNA library for Thellungiella halophila using recombination cloning [J]. J Integr Plant Biol, 2007, 49(9):1313-1319
[19]	方玉达,刘耀光,陈佩度,等. 用温度循环提高DNA连接效率[J]. 农业生物技术学报,1997,7(2): 168,178
[20]	Amtmann A, Bohnert H J, Bressan R A. Abiotic stress and plant genome evolution. search for new models[J]. Plant Physiol ,2005, 138(1):127-130
[21]	Frommer W B, Hummel S, Riesmeier J W. Expression cloning in yeast of a cDNA encoding a broad specificity amino acid permease from Arabidopsis thaliana [J]. Proc Natl Acad Sci USA, 1993, 90(13): 5944-5948

Construction of Stress Induced Full Length cDNA Library of Sedum alfredii and Isolation of Genes Related to Cd-tolerance

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