JVAS announces awards - Four best research articles (one each from basic, production, para-clinical and clinical subjects) and one best short communication will be adjudged for awards each year!!!

Journal of Veterinary and Animal Sciences

Volume: 50 Issue: 2

  • Open Access
  • Research Article

DOUBLE DIGEST RESTRICTION ASSOCIATED DNA SEQUENCING FOR IDENTIFICATION OF SINGLE NUCLEOTIDE POLYMORPHISMS IN GOATS

Shalu Elizabeth Simon1, G.Radhika2, T.V. Aravindakshan3, Marykutty Thomas4 and K. Raji5

1. M.V.Sc. Scholar Department of Animal Genetics and Breeding, College of Veterinary and Animal Sciences, Mannuthy, 2, 4 Assistant Professor 3 Professor and Head 5 Assistant Professor, Department of Physiology, CVAS, Mannuthy

Year: 2019, Page: 108-113,

Received: Sept. 1, 2018 Accepted: Sept. 18, 2018 Published: July 1, 2019

Abstract

Goat rearing is considered as an Animal husbandry activity which promptly supplements economy of the rural poor. India, with its vast animal genetic resources is blessed with 28 goat breeds which vary in different quantitative traits. Malabari and Attappady Black are two native goat breeds of Kerala, which differ significantly in several economic traits like prolificacy, milk production, milk quality and disease resistance. In this study, a novel reduced representation sequencing technique called double digest restriction associated DNA sequencing (ddRADseq) is applied on Malabari and Attappady Black goats, for identification of genetic variants which can be used for exploring many complex economical traits and to serve as markers for selection. DNA samples were isolated from 10 animals of each breed and were pooled into two samples to represent Malabari and Attappady Black goats. These two samples were digested using SphI and MlucI Restriction enzymes which resulted in 26,77,648 reads in Malabari and 17,40,350 reads in Attappady Black goats. A total of 66299, 28859 and 4266 variants were identified in Malabari in read depths of 2, 5 and 10, respectively. In case of Attappady Black, it was 61241, 21230 and 2251. Variants in read depth 10, were considered as high confidence variants. In Read depth 10, Malabari had 4140 SNPs which included 14 missense SNPs, 24 synonymous SNPs, 58 SNPs in untranslated regions and 4044 SNPs in noncoding regions. In Attappady Black groups, 29 missense SNPs, 30 synonymous SNPs, 82 SNPs in untranslated region and 2051 SNPs in non coding regions were present in 2192 high confidence SNPs discovered.

 

Keywords: double digest restriction associated DNA sequencing, Prolificacy, Malabari and Attappady Black goats

References

  1. Brown, J.K., Taggart, J.B., Bekaert, M., Wehner, S., Palaiokostas, C., Setiawan, A.N., Symonds, J.E. and Penman, D.J. 2016. Mapping the sex determination locus in the hapuku (Polyprion oxygeneios) using ddRAD sequencing. BMC genomics. 17: 448-460.
  2. Dimens, P. 2016. Population structure of a migratory small coastal shark, the blacknose shark Carcharhinusacronotus, across cryptic barriers to gene flow. Ph.D. thesis, Texas A&M University-Corpus Christi, 322p
  3. Kai, W., Nomura, K., Fujiwara, A., Nakamura, Y., Yasuike, M., Ojima, N., Masaoka, T., Ozaki, A., Kazeto, Y., Gen, K. and Nagao, J. 2014. A ddRAD-based genetic map and its integration with the genome duplication. BMC genomics. 15: 233- 237.
  4. Kess, T., Gross, J., Harper, F. and Boulding, E.G. 2016. Low-cost ddRAD method of SNP discovery and genotyping applied to the periwinkle Littorina saxatilis. J. Molluscan Stud. 82 : 104-109.
 

  1. Langmead, B., Trapnell, C., Pop, M. and Salzberg, S.L. 2009. Ultrafast and memory-efficient alignment of short DNA sequences to the human genome. Genome Biology. 10: 25-29.
  2. Li, H., Handsaker, B., Wysoker, A., Fennell, T., Ruan, J., Homer, N., Marth, G., Abecasis, G.and         Durbin,  R. 2009.  The  sequence  alignment/map  format  andSAMtools. Bioinformatics. 25: 2078-2079.
  3. Mora-Marquez, F., Garcia-Olivweres, V., Emerson, B.C. and Lopez de Heredia, U., 2017. ddradseqtools: a softwwere package for in silico simulation and testing of double- digest RAD seq experiments. Mol. Ecol. Res. 17: 230- 246
  4. Peterson, B.K., Weber, J.N., Kay, E.H., Fisher, H.S. and Hoekstra, H.E. 2012. Double digest RADseq: an inexpensive method for de novo SNP discovery and genotyping in model and non-model species. PloS one. 7: 35- 44.
  5. Poland, J.A., Brown, P.J., Sorrells, M.E. and Jannink, J.L. 2012. Development of high-density genetic maps for barley and wheat using a novel two-enzyme genotyping-by-sequencing approach. PloS one. 7: 322- 330.
  6. Rasic, G., Filipovic, I., Weeks, A.R. and Hoffmann, A.A. 2014. Genome-wide SNPs lead to strong signals of geographic structure and relatedness patterns in the major arbovirus vector, Aedes aegypti. BMC genomics. 15: 275- 280
  7. Sambrook, J. and Russell, D.W. 2001. Molecular cloning: A laboratory manual. (3rd  Ed.).​​​​​​​Cold spring Harbor Laboratory Press, New York, 1886p.
  8. Yang, C.H., Cheng, Y.H. and Chuang, L.Y. 2010. A natural PCR-RFLP primer design for SNP genotyping using a genetic algorithm. In: The International Multi Conference of Engineers and Computer Scientists. 17 to 19 March, 2010, Hong kong. International Association of Engineers. pp. 124-129.
  9. Yu, H., You, X., Li, J., Liu, H., Meng, Z., Xiao, L., Zhang, H., Lin, H.R., Zhang, Y. and Shi,
  10. 2016. Genome-wide mapping of growth-related quantitative trait loci in orange-spotted grouper (Epinephelus coioides) using double digest restriction-site associated DNA sequencing (ddRADseq). Int J. Mol.Sci. 17: 501- 507
 

  1. Hrdlickova, B., de Almeida, R.C., Borek, Z. and Withoff, S. 2014. Genetic variation in the non-coding genome: Involvement of micro-RNAs and long non-coding RNAs in disease. Biochimicaet Biophysica Acta Molecular Basis of Disease. 1842:1910-1922.

 

 

 

 

Views
74
Downloads
37