Real-time PCR

Age-dependent expression of the BPI gene in Sutai piglets

J. Zhu, Zi, C., Wu, Z. C., Liu, L., Zheng, X. R., Su, X. M., Zhu, G. Q., Huang, X. G., Wu, S. L., and Bao, W. B., Age-dependent expression of the BPI gene in Sutai piglets, vol. 12, pp. 2120-2126, 2013.

We compared and analyzed the expression of the BPI gene of Sutai piglets ranging from newborn to post-weaning days 8, 18, 30, and 35 by the real-time PCR method, in order to determine if it is involved in protection against disease caused by ETEC F18. There was a significant difference between 18 and 35-day expres­sion in the jejunum. There were also significant differences between 35-day expression and expression at the other development stages in the duodenum. There were no significant differences in expression at 8, 18, and 30 days in the jejunum.

TLR4 gene expression in pig populations and its association with resistance to Escherichia coli F18

J. Wang, Pan, Z. Y., Zheng, X. R., Wu, Z. C., Su, X. M., Zhu, G. Q., Huang, X. G., Wu, S. L., and Bao, W. B., TLR4 gene expression in pig populations and its association with resistance to Escherichia coli F18, vol. 12, pp. 2625-2632, 2013.

TLR4 is the main recognition receptor of bacterial lipopolysaccharides, which play an important role in innate and adaptive immunity. We used real-time PCR to analyze the tissue expression profile and differential expression of TLR4 in 4 pig populations (Escherichia coli F18-resistant Sutai, E. coli F18-sensitive Sutai, Large White, Meishan), in order to determine the role that the TLR4 gene plays in resistance to E. coli F18.

Relationships among lipid peroxidation, SOD enzyme activity, and SOD gene expression profile in Lycopersicum esculentum L. exposed to cold stress

S. S. Aydin, Büyük, İ., and Aras, S., Relationships among lipid peroxidation, SOD enzyme activity, and SOD gene expression profile in Lycopersicum esculentum L. exposed to cold stress, vol. 12, pp. 3220-3229, 2013.

The current study was designed to evaluate lipid peroxidation (via malondialdehyde) levels, the superoxide dismutase (SOD) gene expression profile, and SOD enzyme activity in tomato plants (Lycopersicum esculentum L.) subjected to different time periods of cold stress (control, 2, 4, 6, 8, and 10 days). Results revealed that maximum lipid peroxidation occurred in plants exposed to cold stress for 10 days, and SOD enzyme activity gradually increased with increasing exposure to cold stress.

A high-throughput, high-quality plant genomic DNA extraction protocol

H. Li, Li, J., Cong, X. H., Duan, Y. B., Li, L., Wei, P. C., Lu, X. Z., and Yang, J. B., A high-throughput, high-quality plant genomic DNA extraction protocol, vol. 12, pp. 4526-4539, 2013.

The isolation of high-quality genomic DNA (gDNA) is a crucial technique in plant molecular biology. The quality of gDNA determines the reliability of real-time polymerase chain reaction (PCR) analysis. In this paper, we reported a high-quality gDNA extraction protocol optimized for real-time PCR in a variety of plant species. Performed in a 96-well block, our protocol provides high throughput. Without the need for phenol-chloroform and liquid nitrogen or dry ice, our protocol is safer and more cost-efficient than traditional DNA extraction methods.

Expression pattern of the zona pellucida 3 (ZP3) gene during ovarian development and the location of ZP3 protein in oocytes in a natural, wild triploid crucian carp mutant, Carassius auratus var. Pingxiangnensis

J. W. Shi, Sheng, J. Q., Peng, K., Wang, J. H., Yi, W. J., Wu, H. J., Gu, Q., and Hong, Y. J., Expression pattern of the zona pellucida 3 (ZP3) gene during ovarian development and the location of ZP3 protein in oocytes in a natural, wild triploid crucian carp mutant, Carassius auratus var. Pingxiangnensis, vol. 12, pp. 5640-5650, 2013.

Carassius auratus var. Pingxiangnensis (designated CaP), distributed in the Pingxiang region of Jiangxi Province, China, is a natural, wild triploid crucian carp mutant that has two reproductive development modes: gynogenesis and bisexual reproduction. Little information is available about the expression pattern of the zona pellucida 3 (ZP3) gene during ovarian development and the location of the ZP3 protein in oocytes of this fish. In this study, we obtained the full-length cDNA of ZP3 (CaP_ZP3).

Quantitative detection of the rice false smut pathogen Ustilaginoidea virens by real-time PCR

H. Li, Ni, D. H., Duan, Y. B., Chen, Y., Li, J., Song, F. S., Li, L., Wei, P. C., and Yang, J. B., Quantitative detection of the rice false smut pathogen Ustilaginoidea virens by real-time PCR, vol. 12, pp. 6433-6441, 2013.

Rice false smut (RFS) is an important rice disease that is caused by the pathogen Ustilaginoidea virens. In this study, we developed a real-time polymerase chain reaction (PCR) assay to detect U. virens and to estimate the level of disease. The genomic DNAs of U. virens and rice were extracted together from the rice samples. Real-time PCR assays were performed and compared to conventional nested-PCR assays. The real-time PCR assay presented a consistent linearity of the standard curve (R2 = 0.9999).

Interleukin-8 expression associated with canine mammary tumors

D. A. P. C. Zuccari, Castro, R., Gelaleti, G. B., and Mancini, U. M., Interleukin-8 expression associated with canine mammary tumors, vol. 10, pp. 1522-1532, 2011.

The use of prognostic markers for mammary cancer is important for routine diagnosis and research. Interleukin-8 (IL-8) is a chemotactic cytokine, produced by several cell types in response to inflammation. The expression, regulation and function of IL-8 in dogs are little known. Recent studies have associated angiogenesis and inflammatory processes with tumor malignancy. We investigated a possible correlation between IL-8 expression and mammary tumor prognosis in female dogs. IL-8 expression was measured in 50 dogs with mammary neoplasia by immunohistochemistry and real-time PCR.

Differential expression of genes during mastitis in Holstein-Zebu crossbreed dairy cows

I. Fonseca, Antunes, G. R., Paiva, D. S., Lange, C. C., Guimarães, S. E. F., and Martins, M. F., Differential expression of genes during mastitis in Holstein-Zebu crossbreed dairy cows, vol. 10, pp. 1295-1303, 2011.

Among the potential public health problems of animal production, infectious-contagious diseases stand out. Mastitis is among the main diseases affecting dairy cattle. One of the most promising options to reduce the problems caused by this disease, besides proper sanitary and management practices, is selective breeding of resistant animals.

Identification and expression analysis of the Broad-Complex core protein isoform 6 (BR-C Z6) gene in the giant tiger shrimp Penaeus monodon (Penaeidae: Decapoda)

A. Buaklin, Klinbunga, S., and Mensveta, P., Identification and expression analysis of the Broad-Complex core protein isoform 6 (BR-C Z6) gene in the giant tiger shrimp Penaeus monodon (Penaeidae: Decapoda), vol. 10, pp. 2290-2306, 2011.

Broad-Complex (BR-C) is an early ecdysone-responsive gene encoding a family of zinc-finger transcription factors that function during metamorphosis in insects. We identified two full-length cDNAs of BR-C Z6 in the giant tiger shrimp (Penaeus monodon). They were 2422 and 2060 bp in length, containing open reading frames of 1440 and 1443 bp, corresponding to polypeptides of 479 and 480 amino acids, respectively. Tissue distribution analysis indicated that PmBR-C Z6 was abundantly expressed in hemocytes and ovaries in juveniles.

Distribution of HLA-B27 and CYP2D6*4 mutations in the middle Black Sea area (Tokat) of Turkey

S. Sahin, Aydogan, L., Benli, I., and Ozyurt, H., Distribution of HLA-B27 and CYP2D6*4 mutations in the middle Black Sea area (Tokat) of Turkey, vol. 10, pp. 3987-3991, 2011.

We analyzed distribution of HLA-B27 and CYP2D6*4 mutations in 249 patients from Tokat province in Turkey with symptoms of arthritis, sacroiliac, joint and back pain, using a LightCycler 480 II Real-Time PCR thermal cycler. The Genes-4U was applied for studying HLA-B27 mutation, and the Tib-Molbiol commercial kit was used to examine the CYP2D6*4 mutation. Among the 249 patients, 18.5% had the HLA-B27 mutation. The CYP2D6*4 mutation was found in 22.0% (six homozygotes). Ten patients had both mutations. These frequencies are similar to what has been reported from other populations.

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