title: Identification of internal control genes in milk-derived mammary epithelial cells during lactation cycle of Indian zebu cow.
abstract: The present study aims to evaluate the suitability of 10 candidate genes, namely GAPDH, ACTB, RPS15A, RPL4, RPS9, RPS23, HMBS, HPRT1, EEF1A1 and UBI as internal control genes (ICG) to normalize the transcriptional data of mammary epithelial cells (MEC) in Indian cows. A total of 52 MEC samples were isolated from milk of Sahiwal cows (major indigenous dairy breed of India) across different stages of lactation: Early (5-15 days), Peak (30-60 days), Mid (100-140 days) and Late (> 240 days). Three different statistical algorithms: geNorm, Normfinder and BestKeeper were used to assess the suitability of these genes. In geNorm analysis, all the genes exhibited expression stability (M) values below 0.5 with EEF1A1 and RPL4 showing the maximum expression stability. Similar to geNorm, Normfinder also identified EEF1A1 and RPL4 as two of the most stable genes. In Bestkeeper algorithm as well, all the 10 genes showed consistent expression levels. The analysis showed that four genes, that is, EEF1A1, RPL4, GAPDH and ACTB exhibited higher coefficient of correlation to the Bestkeeper index, lower coefficient of variance and standard deviation, indicating their superiority to be used as ICG. The present analysis has provided evidence that RPL4, EEF1A1, GAPDH and ACTB could probably act as most suitable genes for normalizing the transcriptional data of milk-derived mammary epithelial cells of Indian cows.
authors.affiliation: National Bureau of Animal Genetic Resources, Haryana, India.
authors.forename: Pradeep
authors.initials: P
authors.lastname: Jatav
authors.affiliation: National Bureau of Animal Genetic Resources, Haryana, India.
authors.forename: Monika
authors.initials: M
authors.lastname: Sodhi
authors.affiliation: National Bureau of Animal Genetic Resources, Haryana, India.
authors.forename: Ankita
authors.initials: A
authors.lastname: Sharma
authors.affiliation: National Bureau of Animal Genetic Resources, Haryana, India.
authors.forename: Sandeep
authors.initials: S
authors.lastname: Mann
authors.affiliation: National Bureau of Animal Genetic Resources, Haryana, India.
authors.forename: Amit
authors.initials: A
authors.lastname: Kishore
authors.affiliation: National Bureau of Animal Genetic Resources, Haryana, India.
authors.forename: Umesh K
authors.initials: UK
authors.lastname: Shandilya
authors.affiliation: National Dairy Research Institute, Haryana, India.
authors.forename: Ashok K
authors.initials: AK
authors.lastname: Mohanty
authors.affiliation: National Bureau of Animal Genetic Resources, Haryana, India.
authors.forename: Ranjit S
authors.initials: RS
authors.lastname: Kataria
authors.affiliation: National Bureau of Animal Genetic Resources, Haryana, India.
authors.forename: Poonam
authors.initials: P
authors.lastname: Yadav
authors.affiliation: National Bureau of Animal Genetic Resources, Haryana, India.
authors.forename: Preeti
authors.initials: P
authors.lastname: Verma
authors.affiliation: National Dairy Research Institute, Haryana, India.
authors.forename: Surinder
authors.initials: S
authors.lastname: Kumar
authors.affiliation: National Dairy Research Institute, Haryana, India.
authors.forename: Dhruba
authors.initials: D
authors.lastname: Malakar
authors.affiliation: National Bureau of Animal Genetic Resources, Haryana, India.
authors.forename: Manishi
authors.initials: M
authors.lastname: Mukesh
mesh_terms: D000818:Animals; D002417:Cattle / Q000254:growth & development* / Q000502:physiology*; D004847:Epithelial Cells; D005260:Female; D015870:Gene Expression; D006801:Humans; D007774:Lactation; D008321:Mammary Glands, Animal; D008892:Milk / Q000166:cytology*; D060888:Real-Time Polymerase Chain Reaction; D012269:Ribosomal Proteins / Q000235:genetics*; D014158:Transcription, Genetic / Q000235:genetics*
chemical_list: D012269:Ribosomal Proteins
keywords: gene expression; internal control gene; lactation; mammary epithelial cells; qPCR