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Identification and quantitative mRNA analysis of a novel splice variant of GPIHBP1 in dairy cattle.

Yang J, Liu X, Zhang Q, Jiang L - J Anim Sci Biotechnol (2014)

Bottom Line: However, the mRNA structure of the bovine GPIHBP1 gene is not fully determined up to now.With semi-quantitative PCR and quantitative real-time RT-PCR, we found that the mRNA expression of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 in mammary gland of lactating cows were much higher than that in other tissues.Our study reports a novel alternative splicing of GPIHBP1 in bovine for the first time and provide useful information for the further functional analyses of GPIHBP1 in dairy cattle.

View Article: PubMed Central - PubMed

Affiliation: National Engineering Laboratory for Animal Breeding; Key Laboratory of Animal Genetics, Breeding and Reproduction, Ministry of Agriculture of China; College of Animal Science and Technology, China Agricultural University, Beijing, 100193 China.

ABSTRACT

Background: Identification of functional genes affecting milk production traits is very crucial for improving breeding efficiency in dairy cattle. Many potential candidate genes have been identified through our previous genome wide association study (GWAS). Of these, GPIHBP1 is an important novel candidate gene for milk production traits. However, the mRNA structure of the bovine GPIHBP1 gene is not fully determined up to now.

Results: In this study, we identified a novel alternatively splice transcript variant (X5) which leads to a 31 bp insertion in exon 3 and also confirmed the other four existed transcripts (X1, X2, X3 and X4) of the bovine GPIHBP1 gene. We showed that transcript X5 with a 31 bp insertion and transcript X1 with an 8 bp deletion might have tremendous effect on the protein function and structure of GPIHBP1, respectively. With semi-quantitative PCR and quantitative real-time RT-PCR, we found that the mRNA expression of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 in mammary gland of lactating cows were much higher than that in other tissues.

Conclusions: Our study reports a novel alternative splicing of GPIHBP1 in bovine for the first time and provide useful information for the further functional analyses of GPIHBP1 in dairy cattle.

No MeSH data available.


The mRNA expression patterns ofGPIHBP1,GPIHBP1-X1 andGPIHBP1-X5 revealed by RT-PCR. The histograms represent the mRNA expression level of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 in eight tissues of three cows. mRNA expression levels in mammary gland of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 all were highest among 8 tissues. The different capital letters indicated significant differences in the expression among eight tissues at P <0.01.
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Fig5: The mRNA expression patterns ofGPIHBP1,GPIHBP1-X1 andGPIHBP1-X5 revealed by RT-PCR. The histograms represent the mRNA expression level of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 in eight tissues of three cows. mRNA expression levels in mammary gland of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 all were highest among 8 tissues. The different capital letters indicated significant differences in the expression among eight tissues at P <0.01.

Mentions: TaqMan Real-time PCR analysis was conducted to further identify the tissue mRNA expression pattern of bovine GPIHBP1. After normalization with the corresponding mRNA expression level of the housekeeping gene GAPDH, analysis of variance (ANOVA) and multiple comparisons were conducted with R software. We found that mRNA expression level of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 all were significantly different among eight tissues (P < 0.05), in which the mRNA expression levels were significantly higher in mammary gland than other tissues (P < 0.05). And all of GPIHBP1-X1, GPIHBP1-X5 and overall GPIHBP1 had much lower expression level in liver, kidney and muscle (Figure 5).Figure 5


Identification and quantitative mRNA analysis of a novel splice variant of GPIHBP1 in dairy cattle.

Yang J, Liu X, Zhang Q, Jiang L - J Anim Sci Biotechnol (2014)

The mRNA expression patterns ofGPIHBP1,GPIHBP1-X1 andGPIHBP1-X5 revealed by RT-PCR. The histograms represent the mRNA expression level of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 in eight tissues of three cows. mRNA expression levels in mammary gland of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 all were highest among 8 tissues. The different capital letters indicated significant differences in the expression among eight tissues at P <0.01.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4373091&req=5

Fig5: The mRNA expression patterns ofGPIHBP1,GPIHBP1-X1 andGPIHBP1-X5 revealed by RT-PCR. The histograms represent the mRNA expression level of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 in eight tissues of three cows. mRNA expression levels in mammary gland of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 all were highest among 8 tissues. The different capital letters indicated significant differences in the expression among eight tissues at P <0.01.
Mentions: TaqMan Real-time PCR analysis was conducted to further identify the tissue mRNA expression pattern of bovine GPIHBP1. After normalization with the corresponding mRNA expression level of the housekeeping gene GAPDH, analysis of variance (ANOVA) and multiple comparisons were conducted with R software. We found that mRNA expression level of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 all were significantly different among eight tissues (P < 0.05), in which the mRNA expression levels were significantly higher in mammary gland than other tissues (P < 0.05). And all of GPIHBP1-X1, GPIHBP1-X5 and overall GPIHBP1 had much lower expression level in liver, kidney and muscle (Figure 5).Figure 5

Bottom Line: However, the mRNA structure of the bovine GPIHBP1 gene is not fully determined up to now.With semi-quantitative PCR and quantitative real-time RT-PCR, we found that the mRNA expression of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 in mammary gland of lactating cows were much higher than that in other tissues.Our study reports a novel alternative splicing of GPIHBP1 in bovine for the first time and provide useful information for the further functional analyses of GPIHBP1 in dairy cattle.

View Article: PubMed Central - PubMed

Affiliation: National Engineering Laboratory for Animal Breeding; Key Laboratory of Animal Genetics, Breeding and Reproduction, Ministry of Agriculture of China; College of Animal Science and Technology, China Agricultural University, Beijing, 100193 China.

ABSTRACT

Background: Identification of functional genes affecting milk production traits is very crucial for improving breeding efficiency in dairy cattle. Many potential candidate genes have been identified through our previous genome wide association study (GWAS). Of these, GPIHBP1 is an important novel candidate gene for milk production traits. However, the mRNA structure of the bovine GPIHBP1 gene is not fully determined up to now.

Results: In this study, we identified a novel alternatively splice transcript variant (X5) which leads to a 31 bp insertion in exon 3 and also confirmed the other four existed transcripts (X1, X2, X3 and X4) of the bovine GPIHBP1 gene. We showed that transcript X5 with a 31 bp insertion and transcript X1 with an 8 bp deletion might have tremendous effect on the protein function and structure of GPIHBP1, respectively. With semi-quantitative PCR and quantitative real-time RT-PCR, we found that the mRNA expression of GPIHBP1, GPIHBP1-X1 and GPIHBP1-X5 in mammary gland of lactating cows were much higher than that in other tissues.

Conclusions: Our study reports a novel alternative splicing of GPIHBP1 in bovine for the first time and provide useful information for the further functional analyses of GPIHBP1 in dairy cattle.

No MeSH data available.