PCR-SSCP

Associations between polymorphisms of the GFI1B gene and growth traits of indigenous Chinese goats

H. F. Cai, Chen, Z., and Luo, W. X., Associations between polymorphisms of the GFI1B gene and growth traits of indigenous Chinese goats, vol. 13, pp. 872-880, 2014.

This study aimed to investigate polymorphisms of the eighth exon in the GFI1B gene among three indigenous Chinese goat breeds (QianBei Ma goats, GuiZhou white goats, and GuiZhou black goats). Furthermore, association analysis was conducted between these polymorphisms and growth traits. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), direct DNA sequencing, and PCR-restricted fragment length polymorphism (RFLP) were applied to detect polymorphism sites, and a general linear model was used to analyze their association with growth traits.

Ovine prion protein genotype frequencies in northwestern China

C. - L. Zhao, Wu, R., Liu, L., Li, F. - D., Zhang, X. - L., Wang, C., Wang, F., Diao, X. - L., Guan, H. - W., Wang, X., and Zhou, L., Ovine prion protein genotype frequencies in northwestern China, vol. 11, pp. 1671-1681, 2012.

Anti-scrapie breeding programs have been initiated to screen for scrapie-resistant sheep based on ovine prion protein gene (PRNP) genotypes at codons 136, 154 and 171 in many countries, especially European Union member states. However, investigation of sheep PRNP genotypes is limited in China, despite the large number of sheep breeds. We analyzed 432 sheep of five different breeds from farms in northwestern China, using PCR-single-strand conformational polymorphism analysis (PCR-SSCP); the corresponding haplotypes of different PRNP alleles were cloned.

Identification of ANAPC13 gene polymorphisms associated with body measurement traits in Bos taurus

B. J. Jiang, Zhan, X. L., Fu, C. Z., Wang, H. B., Cheng, G., and Zan, L. S., Identification of ANAPC13 gene polymorphisms associated with body measurement traits in Bos taurus, vol. 11, pp. 2862-2870, 2012.

Body measurement traits, influenced by genes and environmental factors, play numerous important roles in the value assessment of productivity and economy. There has been some indication that ANAPC13 influences adult height. We used PCR-SSCP and DNA sequencing technology to identify polymorphisms in the ANAPC13 gene. A polymorphism in intron 1 (A > G at base 17) was identified and an additional polymorphic site (C > T at base 42) was also uncovered, which accompanied the previous polymorphism in more than 98% of the subjects.

Polymorphisms of the myostatin gene (MSTN) and its relationship with growth traits in goat breeds

Z. J. Zhang, Ling, Y. H., Wang, L. J., Hang, Y. F., Guo, X. F., Zhang, Y. H., Ding, J. P., and Zhang, X. R., Polymorphisms of the myostatin gene (MSTN) and its relationship with growth traits in goat breeds, vol. 12. pp. 965-971, 2013.

Mutations in the myostatin (MSTN) gene can inactivate its expression and result in a non-functional protein, which leads to dramatic muscularity and a “double-muscling” phenomenon in many species. Using gene sequencing and polymerase chain reaction-single-strand conformation polymorphism methods, polymorphisms of the MSTN gene were investigated as a candidate marker for growth in 288 goats. The results showed 2 novel single nucleotide polymorphisms: DQ167575 g.197G>A and 345A>T.

Cloning, sequencing, and polymorphisms of the Wistar-Imamichi rat growth hormone gene using PCR-SSCP

M. Q. Liu, Wang, J. G., and Li, X. L., Cloning, sequencing, and polymorphisms of the Wistar-Imamichi rat growth hormone gene using PCR-SSCP, vol. 12, pp. 6203-6211, 2013.

We successfully cloned the Wistar-Imamichi (WI) rat growth hormone gene (GenBank accession: GQ890681), which contained 5 exons and 4 introns. Using the polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) technique, a novel missense substitution single nucleotide polymorphism was identified and tested for Hardy-Weinberg equilibrium with the χ2 test. This gene fragment was investigated in 50 adult rats and 50 neonatal rats by PCR-SSCP.

Novel SNPs in the exon region of bovine DKK4 gene and their association with body measurement traits in Qinchuan cattle

J. B. Gao, Li, Y. K., Yang, N., Ma, X. H., Adoligbe, C., Jiang, B. J., Fu, C. Z., Cheng, G., and Zan, L. S., Novel SNPs in the exon region of bovine DKK4 gene and their association with body measurement traits in Qinchuan cattle, vol. 12, pp. 6664-6672, 2013.

The aim of this study was to determine whether single nucleotide polymorphisms (SNPs) of bovine Dickkopf homolog 4 (DKK4) are associated with body measurement traits in Qinchuan cattle. By using PCR-SSCP technology and DNA sequencing, we discovered 5 DKK4 SNPs in Qingchuan cattle, including -65G>A and -77G>T in the 5ꞌ-untranslated region, 1532C>G and 1533T>C in exon 2, and 2088C>T in exon 3. The sequencing map showed that 1532C>G and 1533T>C were in close linkage disequilibrium and were treated as 1532C>G-1533T>C in this study.

A novel polymorphism of the lactoferrin gene and its association with milk composition and body traits in dairy goats

B. L. Guo, Jiao, Y., He, C., Wei, L. X., Chang, Z. H., Yue, X. P., Lan, X. Y., Chen, H., and Lei, C. Z., A novel polymorphism of the lactoferrin gene and its association with milk composition and body traits in dairy goats, vol. 9, pp. 2199-2206, 2010.

Milk composition and body measurement traits, influenced by genes and environmental factors, play important roles in value assessments of efficiency and productivity in dairy goats. Lactoferrin (LF), involved in the efficient expression of protein in milk, is also an anabolic factor in skeletal tissue and a potent osteoblast survival factor. Therefore, it is an important candidate gene for milk composition and body measurement trait selection in marker-assisted selection.

Polymorphism of somatostatin gene and its association with growth traits in Chinese cattle

L. Gao, Zan, L. S., Wang, H. B., Hao, R. J., and Zhong, X., Polymorphism of somatostatin gene and its association with growth traits in Chinese cattle, vol. 10, pp. 703-711, 2011.

Somatostatins play a crucial role in the regulation of growth and development in vertebrates, especially muscle growth. We assessed the association of somatostatin gene polymorphisms with growth traits by PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism) and DNA sequencing methods in 694 individuals from six Chinese cattle breeds. A novel single nucleotide polymorphism, G126A, was detected, and significant associations were found with body length, body height, hip width, heart girth, and hucklebone width index.

A novel SNP of the C/EBPα gene associated with superior meat quality in indigenous Chinese cattle

H. Wang, Zan, L. S., Wang, H. B., and Song, F. B., A novel SNP of the C/EBPα gene associated with superior meat quality in indigenous Chinese cattle, vol. 10, pp. 2069-2077, 2011.

CCAAT/enhancer-binding protein alpha (C/EBPa) is an essential transcriptional factor regulating the differentiation of adipocytes. We report a novel single nucleotide polymorphism (C271A) of the C/EBPa gene in six indigenous Chinese cattle breeds using PCR-SSCP and DNA sequencing methods. Allele frequencies were investigated and evaluated by the χ2 test in 817 individuals; all populations were found to be in Hardy-Weinberg equilibrium.

Molecular characterization, polymorphism of the ACOX1 gene and association with ultrasound traits in Bos taurus

Y. Jiao, Zan, L. S., Liu, Y. F., and Wang, H. B., Molecular characterization, polymorphism of the ACOX1 gene and association with ultrasound traits in Bos taurus, vol. 10, pp. 1948-1957, 2011.

Acyl-coenzyme A oxidase 1 (ACOX1) is the first enzyme in peroxisomal fatty acid β-oxidation; it is rate-limiting and plays a key role in fatty acid metabolism and fat deposition. ACOX1 is an important candidate gene for meat quality selection through marker-assisted selection. Genomic structural analysis showed that bovine ACOX1 shares 86% identity with human ACOX1. Using PCR-SSCP technology, we discovered a single nucleotide polymorphism (SNP) (A1865C) in exon 13 of the ACOX1 gene.

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