tailieunhanh - báo cáo khoa học: " Identification and evaluation of new reference genes in Gossypium hirsutum for accurate normalization of real-time quantitative RT-PCR data"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Identification and evaluation of new reference genes in Gossypium hirsutum for accurate normalization of real-time quantitative RT-PCR data | Artico et al. BMC Plant Biology 2010 10 49 http 1471-2229 10 49 BMC Plant Biology RESEARCH ARTICLE Open Access Identification and evaluation of new reference genes in Gossypium hirsutum for accurate normalization of real-time quantitative RT-PCR data - A . 11 11 2 2 1 Sinara Artico Sarah M Nardeli Osmundo Brilhante Maria Fatima Grossi-de-Sa Marcio Alves-Ferreira Abstract Background Normalizing through reference genes or housekeeping genes can make more accurate and reliable results from reverse transcription real-time quantitative polymerase chain reaction qPCR . Recent studies have shown that no single housekeeping gene is universal for all experiments. Thus suitable reference genes should be the first step of any qPCR analysis. Only a few studies on the identification of housekeeping gene have been carried on plants. Therefore qPCR studies on important crops such as cotton has been hampered by the lack of suitable reference genes. Results By the use of two distinct algorithms implemented by geNorm and NormFinder we have assessed the gene expression of nine candidate reference genes in cotton GhACT4 GhEF1a5 GhFBX6 GhPP2A1 GhMZA GhPTB GhGAPC2 Gh 3TUB3 and GhUBQ14. The candidate reference genes were evaluated in 23 experimental samples consisting of six distinct plant organs eight stages of flower development four stages of fruit development and in flower verticils. The expression of GhPP2A1 and GhUBQ14 genes were the most stable across all samples and also when distinct plants organs are examined. GhACT4 and GhUBQ14 present more stable expression during flower development GhACT4 and GhFBX6 in the floral verticils and GhMZA and GhPTB during fruit development. Our analysis provided the most suitable combination of reference genes for each experimental set tested as internal control for reliable qPCR data normalization. In addition to illustrate the use of cotton reference genes we checked the expression of two cotton MADS-box genes in distinct .