Quantitative real-time PCR

Developmental changes in the expression of the GLUT2 and GLUT4 genes in the longissimus dorsi muscle of Yorkshire and Tibetan pigs

Y. Liang, Yang, X. M., Gu, Y. R., Tao, X., Zhong, Z. Z., Gong, J. J., Chen, X. H., and Lv, X. B., “Developmental changes in the expression of the GLUT2 and GLUT4 genes in the longissimus dorsi muscle of Yorkshire and Tibetan pigs”, vol. 14, pp. 1287-1292, 2015.

Glucose transporter proteins 2 and 4 (GLUT2 and GLUT4) play important roles in glucose transport and energy metabolism. Changes in the levels of GLUT2 and GLUT4 mRNA were measured in longissimus dorsi muscle from the lean Yorkshire and fat Tibetan pig breeds at six different time points (1, 2, 3, 4, 5, and 6 months) with quantitative real-time polymerase chain reactions. The results showed that GLUT2 and GLUT4 mRNA were abundantly expressed in the longissimus dorsi muscle and that the developmental expression patterns were similar in both breeds.

Molecular cloning and expression analysis of MyD88 in spiny head croaker, Collichthys lucidus

C. Sang, Lin, Y., Jiang, K., Zhang, F., Ma, C., Ma, L., and Song, W., “Molecular cloning and expression analysis of MyD88 in spiny head croaker, Collichthys lucidus”, vol. 14, pp. 4666-4676, 2015.

Myeloid differentiation factor 88 (MyD88) is an important adaptor protein involved in toll-like receptor signaling pathways. In this study, a cDNA library from Collichthys lucidus was constructed using the SMART technique. A complete cDNA sequence showing high identity with the conserved sequence of the MyD88 gene was cloned from the cDNA library using expressed sequence tag analysis and rapid amplification of cDNA ends, and then subjected to further investigation. The full-length cDNA of MyD88 from C.

Association analysis between SNPs in the 5'-flanking region of the chicken GRP78 gene, thermotolerance parameters, and tissue mRNA expression

L. N. Kong, Zhang, D. X., Ji, C. L., Zhang, X. Q., and Luo, Q. B., “Association analysis between SNPs in the 5'-flanking region of the chicken GRP78 gene, thermotolerance parameters, and tissue mRNA expression”, vol. 14, pp. 6110-6123, 2015.

Glucose-regulated protein 78 (GRP78) is a molecular chaperone in the endoplasmic reticulum and can be induced by different kinds of environmental and physiological stress. Thus far, the role of the GRP78 gene in thermotolerance in chickens has not been investigated.

mRNA expression profiles of calmodulin and liver receptor homolog-1 genes in chickens

Z. - C. Zhang, Xiao, L. - H., Wang, Y., Chen, S. - Y., Yang, Z. - Q., Zhao, X. - L., Zhu, Q., and Liu, Y. - P., “mRNA expression profiles of calmodulin and liver receptor homolog-1 genes in chickens”, vol. 11, pp. 3482-3489, 2012.

Calmodulin (CALM), a calcium-binding protein, is expressed in the hypothalamic-pituitary-gonadal axis; it plays a pivotal role in the reproductive system by regulating gonadotropin-releasing hormone signaling. Downstream of hypothalamic-pituitary-gonadal signaling pathways, liver receptor homolog-1 (LRH-1) is involved in female gonadal hormone synthesis. In the chicken, although the two genes are known to be associated with reproductive traits, the interaction between gonadotropins and gonadal steroids remains unclear.

Gene expression in the lignin biosynthesis pathway during soybean seed development

A. Baldoni, Von Pinho, E. V. R., Fernandes, J. S., Abreu, V. M., and Carvalho, M. L. M., “Gene expression in the lignin biosynthesis pathway during soybean seed development”, vol. 12, pp. 2618-2624, 2013.

The study of gene expression in plants is fundamental, and understanding the molecular mechanisms involved in important biological processes, such as biochemical pathways or signaling that are used or manipulated in improvement programs, are key for the production of high-quality soybean seeds. Reports related to gene expression of lignin in seeds are scarce in the literature.

Selection and use of reference genes in mouse mammary glands

L. Q. Han, Yang, G. Y., Zhu, H. S., Wang, Y. Y., Wang, L. F., Guo, Y. J., Lu, W. F., Li, H. J., and Wang, Y. L., “Selection and use of reference genes in mouse mammary glands”, vol. 9, pp. 449-456, 2010.

Obtaining quantitative data concerning gene expression is important for understanding milk synthesis in mammary glands. Quantitative real-time PCR (qRT-PCR) is an efficient tool to calculate gene expression; however, it is necessary to find valid reference genes for normalization of qRT-PCR data. We applied the geNorm software to eight commonly used reference genes to identify the most stable and optimal genes for the mouse mammary gland. Based on this analysis, HPRT, RPL and GAPDH are the most appropriate reference genes for data normalization.

mRNA abundance and expression of SLC27A, ACC, SCD, FADS, LPIN, INSIG, and PPARGC1 gene isoforms in mouse mammary glands during the lactation cycle

L. Q. Han, Li, H. J., Wang, Y. Y., Zhu, H. S., Wang, L. F., Guo, Y. J., Lu, W. F., Wang, Y. L., and Yang, G. Y., “mRNA abundance and expression of SLC27A, ACC, SCD, FADS, LPIN, INSIG, and PPARGC1 gene isoforms in mouse mammary glands during the lactation cycle”, vol. 9, pp. 1250-1257, 2010.

The functions of distinct isoforms of solute carrier family 27 transporters (SLC27A1-6), acetyl-CoA carboxylase (ACACA, ACACB), stearoyl-CoA desaturase (SCD1-4), fatty acid desaturase (FADS1-3), LPIN (LPIN1-3), insulin-induced gene (INSIG1, 2), and peroxisome proliferator-activated receptor gamma coactivator1 (PPARGC1A, B) were studied in the mouse mammary gland from pregnancy to lactation. The relative mRNA abundance and percent change in real-time PCR were determined.

Detection of Salmonella Enteritidis in asymptomatic carrier animals: comparison of quantitative real-time PCR and bacteriological culture methods

B. M. Maciel, Dias, J. C. T., Romano, C. C., Sriranganathan, N., Brendel, M., and Rezende, R. P., “Detection of Salmonella Enteritidis in asymptomatic carrier animals: comparison of quantitative real-time PCR and bacteriological culture methods”, vol. 10, pp. 2578-2588, 2011.

Quantification of Salmonella in asymptomatic carrier animals can be used to assess microbial risk and monitor the level of contamination in domestic animals used for food production. We examined the sensitivity, specificity and accuracy of real-time qPCR, without pre-enrichment or selective enrichment stages, for the quantification of S. enterica serovar Enteritidis in resistant mice, as a model of asymptomatic carrier animal. The results were compared with those obtained by traditional bacteriological culture methods, the gold standard test.

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