辽宁绒山羊DLX3毛囊表达载体的构建及其转染成纤维细胞的研究

Abstract

Abstract: The complete CDS of DLX3 gene was amplified from skin during Liaoning cashmere goat hair follicle growth phase by RT-PCR using one pair of primer which was designed and synthesized according to the DLX3 gene sequence in GenBank(No:XM_005694267.1),inserted into blunt-vector after DNA sequencing verification,it was then sub-cloned into eukaryotic expression vector pIRES2-EGFP.After the restriction enzyme (XhoⅠ/BamHⅠ) digestion of the eukaryotic expression vector and sequencing,the plasmid had been transfected into the goat fibroblasts by Lipofectamine 2000.We also observed the fluorescent under the microscope,examined the transcription vector pIRES2-DLX3-EGFP by RT-PCR and Western blotting.The results showed that we had successfully cloned goat DLX3 gene and constructed eukaryotic expression vector pIRES2-DLX3-EGFP,we could observe the green fluorescent after 24 h transfection of the plasmid.Then, we amplified the transcription product of 909 bp by RT-PCR.The targeted protein 32.87 ku was also detected by Western blotting.This result has paved the way for investigating the influence of DLX3 on hair follicle development and cycling as well as the research of the regulation mechanism of hair follicle growth and development.

Key words: DLX3; cashmere goat; fibroblasts cell; eukaryotic expression vector

CLC Number:

S826

SHU Guo-tao, YAO Na, DONG Kun-zhe, KANG Ye, PU Ya-bin, ZHAO Qian-jun, HE Xiao-hong, MA Yue-hui. Construction of Hair Follicle Expression Vector of DLX3 and its Transfection into Liaoning Cashmere Goat Fibroblasts Cell[J]. , 2014, 41(11): 29-34.

References

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Construction of Hair Follicle Expression Vector of DLX3 and its Transfection into Liaoning Cashmere Goat Fibroblasts Cell

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