Construction of subtracted cDNA library of Malus xiaojinensis is the base of screening and cloning genes under Fe-deficiency stress. Using LD-PCR technique, cDNA was synthesized from total RNA of Malus xiaojinensis roots treated under Fe-deficiency stress(EDTA-Fe2＋，4μmol/L)which was used as tester and Fe-sufficiency(EDTA-Fe2＋，40μmol/L)as driver. A subtracted cDNA Library was constructed using suppression subtractive hybridization(SSH). After two hybridizations and two suppression PCRs, the amplified library contained 1200 positive clones. Plasmid inserts were PCR amplified and showed 300~800bp inserts. There is the same result with RsaⅠdigestion of plasmid
Zhang Yugang, Xu Xuefeng, Li Tianzhong Han Zhenhai,
. Construction of Subtracted cDNA Library of Malus xiaojinensis with Suppression Subtractive Hybridization and Their Quality Analysis[J]. Chinese Agricultural Science Bulletin, 2005
, 21(3)
: 77
-77
.
DOI: 10.11924/j.issn.1000-6850.050377
1Thoiron S, Pascal N, Briat JF. Impact of iron deficiency and iron Fe-suppy during the early stages os vegetative development in maize(Zea mays L.). Plant Cell and Environment, 1997,20:1051~1060
2Han Z H, Xu X F. Current status and the existed problems in research of iron efficient fruit genotypes. Annual Review of Horticultural Science,1995,1:1~16
3刘彦华.苹果铁高效基因型生物技术的研究——根总RNA提取方法的比较及mRNA的纯化：[硕士学位论文].北京:中国农业大学,1999.6~9
4Diatchenlo L, Chris Lau Y F, Campbell A P, et al. Suppression Subtractive Hybridization:a method for generating differentially regulated or tissue-specific cDNA probes and libraries.Proc Natl Acad Sci USA,1996,93: 6025~6030
5顾克余,翟虎渠.抑制性消减杂技术及其在基因克隆上的研究.生物技术通报,1999,2:13~16
6刘军,袁自强,刘建东,钱晓茵,钱旻,杨金水.应用抑制差减杂交法分离水稻幼穗发育早期特异表达的基因.科学通报,2000,45(13):1392~1397
7高鹏.玉米幼苗淹水诱导表达基因的克隆及其功能分析:[博士学位论文].北京:中国农业大学,2003.59~68
8郑军.玉米幼苗水分胁迫相关基因的克隆与分析:[博士学位论文].北京:中国农业大学,2004.44~60
