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Molecular cloning and expression pattern of duck <i style="mso-bidi-font-style:normal">Six1</i> and its preliminary functional analysis in myoblasts transfected with eukaryotic expression vector

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Title Molecular cloning and expression pattern of duck Six1 and its preliminary functional analysis in myoblasts transfected with eukaryotic expression vector
 
Creator Wang, Haohan
Jin, Haibo
Liu, Hehe
Sun, Lingli
Li, Xinxin
Yang, Chao
Zhang, Rongping
Li, Liang
Wang, Jiwen
 
Subject Duck Six1
Expression
Molecular cloning
Myoblast
pEGFP-duSix1
 
Description 271-281
Skeletal muscle development
is regulated by Six1, an important
myogenic transcription factor. However, the functional analysis of duck Six1 has not been reported. Here, we cloned the coding domain sequence (CDS) region of the duck Six1 gene using RT-PCR and RACE methods. Bioinformatics analysis revealed
that duck Six1 CDS region comprised of 849 bp and encoded 282
amino acids and had a high degree of homology with other
species, suggesting that the functions of duck Six1 gene are conserved among other animals. Real-time
PCR used to determine the mRNA expression profiles of duck Six1 in different tissues and different
developmental stages showed that Six1
was highly expressed in skeletal muscle and the embryonic stage. Furthermore, the
eukaryotic expression
vector pEGFP-duSix1
was constructed and transfected into the duck myoblasts; the MTT assay revealed
an obvious increase of cell proliferation after transfection. The expression
profiles of Six1, Myf5 and MyoD showed that their expression levels
were significantly increased. These results together suggested that
pEGFP-duSix1 vector was constructed successfully and overexpression of duck Six1 in the myoblasts could promote cell
proliferation activity and significant up-regulate expression of Myf5 and MyoD.
 
Date 2014-09-03T04:17:02Z
2014-09-03T04:17:02Z
2014-08
 
Type Article
 
Identifier 0975-0959 (Online); 0301-1208 (Print)
http://hdl.handle.net/123456789/29321
 
Language en_US
 
Rights CC Attribution-Noncommercial-No Derivative Works 2.5 India
 
Publisher NISCAIR-CSIR, India
 
Source IJBB Vol.51(4) [August 2014]