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Recombinant Goat VEGF164 Increases Hair Growth by Painting Process on the Skin of Shaved Mouse.

Bao W, Yin J, Liang Y, Guo Z, Wang Y, Liu D, Wang X, Wang Z - Asian-australas. J. Anim. Sci. (2014)

Bottom Line: The expression of recombinant 6×his-gVEGF164 protein was induced by 0.5 mM isopropyl thio-β-D-galactoside at 32°C.The rgVEGF164 was smeared onto the dorsal area of a shaved mouse, and we noted that hair regrowth in this area was faster than in the control group.Thus, rgVEGF164 increases hair growth in mice.

View Article: PubMed Central - PubMed

ABSTRACT
To detect goat vascular endothelial growth factor (VEGF)-mediated regrowth of hair, full-length VEGF164 cDNA was cloned from Inner Mongolia cashmere goat (Capra hircus) into the pET-his prokaryotic expression vector, and the recombinant plasmid was transferred into E. coli BL21 cells. The expression of recombinant 6×his-gVEGF164 protein was induced by 0.5 mM isopropyl thio-β-D-galactoside at 32°C. Recombinant goat VEGF164 (rgVEGF164) was purified and identi ed by western blot using monoclonal anti-his and anti-VEGF antibodies. The rgVEGF164 was smeared onto the dorsal area of a shaved mouse, and we noted that hair regrowth in this area was faster than in the control group. Thus, rgVEGF164 increases hair growth in mice.

No MeSH data available.


Analysis of the expression of rgVEGF164 in E. coli BL21 (DE3). (A) Expression of rgVEGF164 analyzed by 15% SDS-PAGE. Lane 1, lysates of E. coli BL21 (DE3) cells before induction; lane 2, lysates of BL21 cells harboring pET-His before induction; lane 3, lysates of BL21 cells harboring pET-gVEGF164 before induction; lane 4, lysates of BL21 cells after induction; lane 5, BL21 lysates harboring pET-His after induction; lane 6, lysates of BL21 cells harboring pET-gVEGF164 after induction; M, low protein molecular weight marker (D530S, Takara Co. Ltd., Dalian, China). The arrowhead indicates the target protein. (B) Purification and identification of rgVEGF164. Lane 1, purified VEGF164; M, unstained protein molecular weight marker (SM0431, Thermo Fisher Sientific Anatomical Pathology, Fremont, CA, USA). VEGF, vascular endothelial growth factor; rgVEGF164, recombinant goat VEFG164; SDS-PAGE, Sodium dodecyl sulfate-polyacrylamide gelelectrophoresis, gVEGF164, goat VEGF164.
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f2-ajas-27-9-1355: Analysis of the expression of rgVEGF164 in E. coli BL21 (DE3). (A) Expression of rgVEGF164 analyzed by 15% SDS-PAGE. Lane 1, lysates of E. coli BL21 (DE3) cells before induction; lane 2, lysates of BL21 cells harboring pET-His before induction; lane 3, lysates of BL21 cells harboring pET-gVEGF164 before induction; lane 4, lysates of BL21 cells after induction; lane 5, BL21 lysates harboring pET-His after induction; lane 6, lysates of BL21 cells harboring pET-gVEGF164 after induction; M, low protein molecular weight marker (D530S, Takara Co. Ltd., Dalian, China). The arrowhead indicates the target protein. (B) Purification and identification of rgVEGF164. Lane 1, purified VEGF164; M, unstained protein molecular weight marker (SM0431, Thermo Fisher Sientific Anatomical Pathology, Fremont, CA, USA). VEGF, vascular endothelial growth factor; rgVEGF164, recombinant goat VEFG164; SDS-PAGE, Sodium dodecyl sulfate-polyacrylamide gelelectrophoresis, gVEGF164, goat VEGF164.

Mentions: Goat VEGF164 was expressed as 6×his-gVEGF164 recombinant protein (rgVEGF164) after induction in E. coli BL21 (DE3) cells that were transformed with pET-gVEGF164. We determined the optimal culture conditions for the rgVEGF164 protein as follows: 0.5 mM IPTG with a 5-h induction at 0.6 of the OD600 value at 32°C. The supernatants of the protein lysates from E. coli BL21 (DE3) cells, the bacterial cells transformed with pET-his and transformed with pET-gVEGF164 were electrophoresed on 15% (w/v) SDS-polyacrylamide gels, and rgVEGF164 was detected by SDS-PAGE (Figure 2A).


Recombinant Goat VEGF164 Increases Hair Growth by Painting Process on the Skin of Shaved Mouse.

Bao W, Yin J, Liang Y, Guo Z, Wang Y, Liu D, Wang X, Wang Z - Asian-australas. J. Anim. Sci. (2014)

Analysis of the expression of rgVEGF164 in E. coli BL21 (DE3). (A) Expression of rgVEGF164 analyzed by 15% SDS-PAGE. Lane 1, lysates of E. coli BL21 (DE3) cells before induction; lane 2, lysates of BL21 cells harboring pET-His before induction; lane 3, lysates of BL21 cells harboring pET-gVEGF164 before induction; lane 4, lysates of BL21 cells after induction; lane 5, BL21 lysates harboring pET-His after induction; lane 6, lysates of BL21 cells harboring pET-gVEGF164 after induction; M, low protein molecular weight marker (D530S, Takara Co. Ltd., Dalian, China). The arrowhead indicates the target protein. (B) Purification and identification of rgVEGF164. Lane 1, purified VEGF164; M, unstained protein molecular weight marker (SM0431, Thermo Fisher Sientific Anatomical Pathology, Fremont, CA, USA). VEGF, vascular endothelial growth factor; rgVEGF164, recombinant goat VEFG164; SDS-PAGE, Sodium dodecyl sulfate-polyacrylamide gelelectrophoresis, gVEGF164, goat VEGF164.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4150203&req=5

f2-ajas-27-9-1355: Analysis of the expression of rgVEGF164 in E. coli BL21 (DE3). (A) Expression of rgVEGF164 analyzed by 15% SDS-PAGE. Lane 1, lysates of E. coli BL21 (DE3) cells before induction; lane 2, lysates of BL21 cells harboring pET-His before induction; lane 3, lysates of BL21 cells harboring pET-gVEGF164 before induction; lane 4, lysates of BL21 cells after induction; lane 5, BL21 lysates harboring pET-His after induction; lane 6, lysates of BL21 cells harboring pET-gVEGF164 after induction; M, low protein molecular weight marker (D530S, Takara Co. Ltd., Dalian, China). The arrowhead indicates the target protein. (B) Purification and identification of rgVEGF164. Lane 1, purified VEGF164; M, unstained protein molecular weight marker (SM0431, Thermo Fisher Sientific Anatomical Pathology, Fremont, CA, USA). VEGF, vascular endothelial growth factor; rgVEGF164, recombinant goat VEFG164; SDS-PAGE, Sodium dodecyl sulfate-polyacrylamide gelelectrophoresis, gVEGF164, goat VEGF164.
Mentions: Goat VEGF164 was expressed as 6×his-gVEGF164 recombinant protein (rgVEGF164) after induction in E. coli BL21 (DE3) cells that were transformed with pET-gVEGF164. We determined the optimal culture conditions for the rgVEGF164 protein as follows: 0.5 mM IPTG with a 5-h induction at 0.6 of the OD600 value at 32°C. The supernatants of the protein lysates from E. coli BL21 (DE3) cells, the bacterial cells transformed with pET-his and transformed with pET-gVEGF164 were electrophoresed on 15% (w/v) SDS-polyacrylamide gels, and rgVEGF164 was detected by SDS-PAGE (Figure 2A).

Bottom Line: The expression of recombinant 6×his-gVEGF164 protein was induced by 0.5 mM isopropyl thio-β-D-galactoside at 32°C.The rgVEGF164 was smeared onto the dorsal area of a shaved mouse, and we noted that hair regrowth in this area was faster than in the control group.Thus, rgVEGF164 increases hair growth in mice.

View Article: PubMed Central - PubMed

ABSTRACT
To detect goat vascular endothelial growth factor (VEGF)-mediated regrowth of hair, full-length VEGF164 cDNA was cloned from Inner Mongolia cashmere goat (Capra hircus) into the pET-his prokaryotic expression vector, and the recombinant plasmid was transferred into E. coli BL21 cells. The expression of recombinant 6×his-gVEGF164 protein was induced by 0.5 mM isopropyl thio-β-D-galactoside at 32°C. Recombinant goat VEGF164 (rgVEGF164) was purified and identi ed by western blot using monoclonal anti-his and anti-VEGF antibodies. The rgVEGF164 was smeared onto the dorsal area of a shaved mouse, and we noted that hair regrowth in this area was faster than in the control group. Thus, rgVEGF164 increases hair growth in mice.

No MeSH data available.