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| Construction of Eukaryotic Expression Vector of MC1R Gene and Its Expression in Hair Follicle Stem Cells of Alpaca (Lama pacos) |
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Abstract Melanocortin-1 receptor (MC1R) gene may play important roles in the regulation process of mammalian coat color, but the effects of exogenous MC1R gene on the micropht-halmiaassocitated transcription factor (MITF) and tyrosinase (TYR) genes in alpaca (Lama pacos) are less studied. To explore effect of MC1R gene on the expression of MITF and TYR, eukaryotic expression vector of MC1R gene pcDNA3.1(+)-MC1R was constructed and transfected into alpaca hair follicle stem cells (HFSCs). The relative expression of MC1R, MITF and TYR mRNA were detected by qRT-PCR, and MC1R protein was located by immunocytochemistry. qRT-PCR results showed that the relative expression level of MC1R was significantly increased (P＜0.05) in the group transfected with pcDNA3.1(+)-MC1R (P＜0.05). There were 2.15 and 2.26 times than that in the group of pcDNA3.1(+) and the control group. MITF and TYR gene also had higher expression, but not significant (P＞0.05). Meanwhile immunocytochemistry results showed that MC1R protein had a positive expression in HFSCs. The results showed that the MC1R gene excessively expressed in alpaca HFSCs, and indirectly up-regulated the expression of TYR and MITF. This study provides scientific basis for further study on the expression mechanism of MC1R in the alpaca HFSCs.
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Received: 21 November 2016
Published: 31 March 2017
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