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Scientia Silvae Sinicae ›› 2006, Vol. 42 ›› Issue (6): 32-36.doi: 10.11707/j.1001-7488.20060606

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Isolation of Differentially Expressed Genes in Latex of Hevea brasiliensis by Suppressive Subtractive Hybridization

Deng Liuhong1,Luo Mingwu2,Zeng Huicai1,Yang Weifan1,Zhang Chunfa1   

  1. 1.State Key Laboratory of Tropical Crop Biotechnology, Chinese Academy of Tropical Agriculture Sciences Haikou 571101;2.South China University of Tropical Agriculture Danzhou 571737
  • Received:2004-10-29 Revised:1900-01-01 Online:2006-06-25 Published:2006-06-25

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Abstract:

A subtracted cDNA library was constructed via suppressive subtractive hybridization(SSH), in which the latex Poly(A+)RNA isolated from regularly tapped Hevea brasiliensis trees was utilized as tester while driver is Poly(A+)RNA derived from negative latex-producing leaf tissues. Total 79 positive clones were identified from screening of the subtracted library by PCR and Reverse Northern Dot-Blot, part of them were further verified by RT_PCR and Northern Blot.18 out of these 79 confirmed clones were randomly selected for sequencing analyses, of which the results were aligned with GenBank? databases via BLASTN and BLASTX programs. It is shown that 5 of 18 candidates were identity with known genes in H. brasiliensis which exerted a positive role in rubber synthesis, 2 showed significant similarity to cis-prenyltransferase of H. brasiliensis, 11 were novel genes in H. brasiliensis.

Key words: Hevea brasiliensis, suppressive subtractive hybridization(SSH), latex, Reverse Northern Dot_Blot