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Construction and characterization of normalized cDNA library of maize inbred MO17 from multiple tissues and developmental stages

  • Genomics. Transcriptomics. Proteomics
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Abstract

A comprehensive complementary DNA (cDNA) library is a valuable resource for functional genomics. In this study, we set up a normalized cDNA library of Mo17 (MONL) by saturation hybridization with genomic DNA, which contained expressed genes of eight tissues and organs from inbred Mo17 of maize (Zea mays L.). In this library, the insert sizes range from 0.4 kb to 4 kb and the average size is 1.18 kb. 10 830 clones were spotted on nylon membrane to make a cDNA microarray. Randomly picked 300 clones from the cDNA library were sequenced. The cDNA microarray was hybridized with pooled tissue mRNA probes or housekeeping gene cDNA probes. The results showed the normalized cDNA library comprehensively includes tissue-specific genes in which 71% are unique ESTs (expressed sequence tags) based on the 300 sequences analyzed. Using the BLAST program to compare the sequences against online nucleotide databases, 88% sequences were found in ZmDB or NCBI, and 12% sequences were not found in existing nucleotide databases. More than 73% sequences are of unknown function. The library could be extensively used in developing DNA markers, sequencing ESTs, mining new genes, identifying positional cloning and candidate genes, and developing microarrays in maize genomics research.

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Abbreviations

C o t :

initial concentration of DNA (mol of nucleotide per liter) × time (sec)

TUs:

Tentative Unigene Sequences

ESTs:

Expressed Sequence Tags

IPTG:

isopropylthio-β-D-galactoside

X-gal:

5-bromo-4-chloro-3-indolyl-β-D-galactoside

MONL:

Mo17 Normalized cDNA Library

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Authors and Affiliations

  1. National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, 430070, China

    Z. X. Zhang, F. D. Zhang, W. H. Tang, Y. J. Pi & Y. L. Zheng

  2. College of Life Science, Yangtze University, Jingzhou, 434025, China

    Z. X. Zhang

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  2. F. D. Zhang
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  3. W. H. Tang
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__________

From Molekulyarnaya Biologiya, Vol. 39, No. 2, 2005, pp. 198–206.

Original English Text Copyright © 2005 by Z. Zhang, F. Zhang, Tang, Pi, Zheng.

This article was submitted by the authors in English.

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Zhang, Z.X., Zhang, F.D., Tang, W.H. et al. Construction and characterization of normalized cDNA library of maize inbred MO17 from multiple tissues and developmental stages. Mol Biol 39, 177–184 (2005). https://doi.org/10.1007/s11008-005-0026-8

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  • DOI: https://doi.org/10.1007/s11008-005-0026-8

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