Lipoprotein Lipase, Tissue Expression and Effects on Genes Related to Fatty Acid Synthesis in Goat Mammary Epithelial Cells
Abstract
:1. Introduction
2. Results and Discussion
2.1. Cloning and Sequence Analysis of Lipoprotein Lipase (LPL)
2.2. Phylogenetic Analysis of LPL Protein
2.3. The Expression Profiles of LPL in 10 Tissues and Different Stages of Lactation
2.4. Effects of LPL Knockdown and Orlistat on mRNA Expression of Genes Related to Milk Fat Synthesis in Goat Mammary Epithelial Cells (GMEC)
3. Experimental Section
3.1. Animals and Samples Collection
3.2. Cloning, Sequencing Analysis and Tissue Expression of LPL
3.2.1. Total RNA Extraction, cDNA Synthesis and Quantification
3.2.2. Full-Length Cloning of LPL cDNA
3.2.3. Bioinformatics Analysis of LPL
3.3. Expression and Statistical Analysis
3.4. Effects of Orlistat (Tetrahydrolipstain, THL) and LPL Knockdown on mRNA Expression of LPL and Genes Related to Milk Fat Synthesis in GMEC
3.4.1. Adenovirus Establishment
3.4.2. Cell Culture and Treatments
4. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Abbreviation
ACACA | acetyl-coenzyme A carboxylase alpha |
ANOVA | analysis of variance |
bp | base pair(s) |
BLAST | basic local alignment search tool |
bMEC | bovine mammary epithelial cells |
cDNA | DNA complimentary to RNA |
CDS | coding sequence |
CTR | control |
DMEM | Dulbecco’s modified Eagle medium |
EC | enzyme class |
EGF | epidermal growth factor |
FASN | fatty acid synthase |
FBS | fetal bovine serum |
FFA | free fatty acids |
GAPDH | glyceraldehyde-3-phosphate dehydrogenase |
GMEC | goat mammary epithelial cells |
FFAR3 | free fatty acid receptor 3 |
LIPE | lipase, hormone-sensitive |
kDa | kilo Dalton(s) |
LCFA | long chain fatty acid |
LPL | Lipoprotein lipase |
MG | mammary gland |
MOI | multiplicity of infection |
MW | molecular weight |
NC | negative control |
NCBI | national center for biotechnology information |
NEFA | non-esterified fatty acids |
NJ | neighbor-joining |
nt | nucleotide |
ORF | open reading frame |
PBS | phosphate buffered saline |
pI | isoelectric point |
PPARG | peroxisome proliferator-activated receptor-γ |
RACE | rapid amplification of cDNA ends |
RPS9 | ribosomal protein S9 |
RT-PCR | reverse transcription polymerase chain reaction |
SCFAs | short-chain fatty acids |
SD | standard deviation |
shRNA | short hairpin RNA |
SREBF1 | sterol regulatory element binding factor1 |
TAG | triacylgleryerol |
TG | triglyceride |
UTR | un-translated region(s) |
UXT | Ubiquitously expressed transcript protein |
VLDL | very-low density lipoproteins |
Conflicts of Interest
References
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Zhao, W.-S.; Hu, S.-L.; Yu, K.; Wang, H.; Wang, W.; Loor, J.; Luo, J. Lipoprotein Lipase, Tissue Expression and Effects on Genes Related to Fatty Acid Synthesis in Goat Mammary Epithelial Cells. Int. J. Mol. Sci. 2014, 15, 22757-22771. https://doi.org/10.3390/ijms151222757
Zhao W-S, Hu S-L, Yu K, Wang H, Wang W, Loor J, Luo J. Lipoprotein Lipase, Tissue Expression and Effects on Genes Related to Fatty Acid Synthesis in Goat Mammary Epithelial Cells. International Journal of Molecular Sciences. 2014; 15(12):22757-22771. https://doi.org/10.3390/ijms151222757
Chicago/Turabian StyleZhao, Wang-Sheng, Shi-Liang Hu, Kang Yu, Hui Wang, Wei Wang, Juan Loor, and Jun Luo. 2014. "Lipoprotein Lipase, Tissue Expression and Effects on Genes Related to Fatty Acid Synthesis in Goat Mammary Epithelial Cells" International Journal of Molecular Sciences 15, no. 12: 22757-22771. https://doi.org/10.3390/ijms151222757