Quantitative real-time PCR

Characterization, molecular cloning, and expression analysis of Ecsit in the spinyhead croaker, Collichthys lucidus

W. Song, Jiang, K. J., Zhang, F. Y., Wang, J., Ma, L. B., Song, W., Jiang, K. J., Zhang, F. Y., Wang, J., Ma, L. B., Song, W., Jiang, K. J., Zhang, F. Y., Wang, J., and Ma, L. B., Characterization, molecular cloning, and expression analysis of Ecsit in the spinyhead croaker, Collichthys lucidus, vol. 15, p. -, 2016.

Evolutionarily conserved signaling intermediate in Toll pathways (Ecsit) is reported to play an essential role in innate immunity, embryogenesis, and assembly or stability of the mitochondrial complex I. In this study, the full-length cDNA of Ecsit was cloned from the spinyhead croaker Collichthys lucidus based on the expressed sequence tags from our cDNA library constructed using the SMART technique.

Characterization of hemocyanin from the mud crab Scylla paramamosain and its expression analysis in different tissues, at various stages, and under Vibrio parahaemolyticus infection

J. Wang, Zhang, F. Y., Song, W., Fang, Y. B., Hu, J. H., Zhao, M., Jiang, K. J., and Ma, L. B., Characterization of hemocyanin from the mud crab Scylla paramamosain and its expression analysis in different tissues, at various stages, and under Vibrio parahaemolyticus infection, vol. 14, pp. 16639-16651, 2015.

Hemocyanin is an important respiratory protein in many arthropod and mollusk species. Here, four cDNAs (SpHc1, SpHc2, SpHc3, and SpHc4), encoding distinct hemocyanin subunits from Scylla paramamosain were cloned using EST analyses and the rapid amplification of cDNA ends. The four full-length cDNA fragments (SpHc1-4) were 2281, 2002, 2184, and 2069 bp, respectively, and they encoded four putative proteins (570-676 amino acids) with a molecular mass of ~65.0-76.8 kDa.

Characterization and expression analysis of the prophenoloxidase activating factor from the mud crab Scylla paramamosain

J. Wang, Jiang, K. J., Zhang, F. Y., Song, W., Zhao, M., Wei, H. Q., Meng, Y. Y., and Ma, L. B., Characterization and expression analysis of the prophenoloxidase activating factor from the mud crab Scylla paramamosain, vol. 14, pp. 8847-8860, 2015.

Prophenoloxidase activating factors (PPAFs) are a group of clip domain serine proteinases that can convert prophenoloxidase (pro-PO) to the active form of phenoloxidase (PO), causing melanization of pathogens. Here, two full-length PPAF cDNAs from Scylla paramamosain (SpPPAF1 and SpPPAF2) were cloned and characterized. The full-length SpPPAF1 cDNA was 1677 bp in length, including a 5'-untranslated region (UTR) of 52 bp, an open reading frame (ORF) of 1131 bp coding for a polypeptide of 376 amino acids, and a 3'-UTR of 494 bp.

Identification and evaluation of endogenous control genes for use in quantitative RT-PCR during wheat (Triticum aestivum L.) grain filling

D. Wu, Dong, J., Yao, Y. J., Zhao, W. C., and Gao, X., Identification and evaluation of endogenous control genes for use in quantitative RT-PCR during wheat (Triticum aestivum L.) grain filling, vol. 14, pp. 10530-10542, 2015.

The use of appropriate reference genes is essential for the generation of accurate and biologically meaningful results from quantitative real-time PCR (qRT-PCR) analysis. However, studies have found that the expression of most commonly used reference genes is not always independent of the tissues, treatments, or developmental stages studied. geNormPlus, NormFinder, and BestKeeper, were applied and the expression stability of nine candidate genes was evaluated in different data sets during wheat grain development.

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