scholarly journals Recombinant human fibroblast growth factor-18 (rhFG18) promotes bovine articular chondrocyte proliferation and cartilage matrix production in vitro

2012 ◽  
Vol 20 ◽  
pp. S135 ◽  
Author(s):  
A. Gigout ◽  
D. Werkmann ◽  
S. Lindemann ◽  
K. Kleinschmidt ◽  
H. Guehring
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Human Fibroblast ◽  
Cartilage Matrix ◽  
Fibroblast Growth ◽  
Articular Chondrocyte ◽  
Chondrocyte Proliferation ◽  
Matrix Production ◽  
And Cartilage

scholarly journals A New 34-Kilodalton Isoform of Human Fibroblast Growth Factor 2 Is Cap Dependently Synthesized by Using a Non-AUG Start Codon and Behaves as a Survival Factor

1999 ◽  
Vol 19 (1) ◽  
pp. 505-514 ◽  
Author(s):  
Emmanuelle Arnaud ◽  
Christian Touriol ◽  
Christel Boutonnet ◽  
Marie-Claire Gensac ◽  
Stéphan Vagner ◽  
...  
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Human Fibroblast ◽  
The Other ◽  
In Vitro Translation ◽  
Start Codon ◽  
Fibroblast Growth Factor 2 ◽  
Initiation Codon ◽  
Fibroblast Growth

ABSTRACT Four isoforms of human fibroblast growth factor 2 (FGF-2) result from alternative initiations of translation at three CUG start codons and one AUG start codon. Here we characterize a new 34-kDa FGF-2 isoform whose expression is initiated at a fifth initiation codon. This 34-kDa FGF-2 was identified in HeLa cells by using an N-terminal directed antibody. Its initiation codon was identified by site-directed mutagenesis as being a CUG codon located at 86 nucleotides (nt) from the FGF-2 mRNA 5′ end. Both in vitro translation and COS-7 cell transfection using bicistronic RNAs demonstrated that the 34-kDa FGF-2 was exclusively expressed in a cap-dependent manner. This contrasted with the expression of the other FGF-2 isoforms of 18, 22, 22.5, and 24 kDa, which is controlled by an internal ribosome entry site (IRES). Strikingly, expression of the other FGF-2 isoforms became partly cap dependent in vitro in the presence of the 5,823-nt-long 3′ untranslated region of FGF-2 mRNA. Thus, the FGF-2 mRNA can be translated both by cap-dependent and IRES-driven mechanisms, the balance between these two mechanisms modulating the ratio of the different FGF-2 isoforms. The function of the new FGF-2 was also investigated. We found that the 34-kDa FGF-2, in contrast to the other isoforms, permitted NIH 3T3 cell survival in low-serum conditions. A new arginine-rich nuclear localization sequence (NLS) in the N-terminal region of the 34-kDa FGF-2 was characterized and found to be similar to the NLS of human immunodeficiency virus type 1 Rev protein. These data suggest that the function of the 34-kDa FGF-2 is mediated by nuclear targets.

scholarly journals Novel Method of Cell-Free In Vitro Synthesis of the Human Fibroblast Growth Factor 1 Gene

2010 ◽  
Vol 2010 ◽  
pp. 1-5
Author(s):  
Peijun Zuo ◽  
A. Bakr M. Rabie
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Human Fibroblast ◽  
Recombinant Dna ◽  
Fibroblast Growth ◽  
In Vitro Synthesis ◽  
Dna Oligonucleotides ◽  
Conventional Procedure ◽  
New Genes

Recombinant DNA projects generally involve cell-based gene cloning. However, because template DNA is not always readily available, in vitro chemical synthesis of complete genes from DNA oligonucleotides is becoming the preferred method for cloning. This article describes a new, rapid procedure based onTaqpolymerase for the precise assembly of DNA oligonucleotides to yield the complete human fibroblast growth factor 1 (FGF1) gene, which is 468 bp long and has a G+C content of 51.5%. The new method involved two steps: (1) the design of the DNA oligonucleotides to be assembled and (2) the assembly of multiple oligonucleotides by PCR to generate the whole FGF1 gene. The procedure lasted a total of only 2 days, compared with 2 weeks for the conventional procedure. This method of gene synthesis is expected to facilitate various kinds of complex genetic engineering projects that require rapid gene amplification, such as cell-free whole-DNA library construction, as well as the construction of new genes or genes that contain any mutation, restriction site, or DNA tag.

scholarly journals Biosynthesis of human fibroblast growth factor-5.

1991 ◽  
Vol 11 (4) ◽  
pp. 1840-1845 ◽  
Author(s):  
B Bates ◽  
J Hardin ◽  
X Zhan ◽  
K Drickamer ◽  
M Goldfarb
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Human Fibroblast ◽  
Point Mutations ◽  
Human Tumor ◽  
Upstream Open Reading Frame ◽  
Fibroblast Growth ◽  
3T3 Cells ◽  
Reading Frame

We have analyzed the biosynthesis of human fibroblast growth factor-5 (FGF-5) at the translational and posttranslational levels. FGF-5 RNA synthesized in vitro can be translated in rabbit reticulocyte lysates to yield a 29,500-Da protein, which is consistent with the molecular weight predicted from the coding sequence. The efficiency of FGF-5 translation is dramatically enhanced if an upstream open reading frame (ORF-1) in the RNA is deleted or if both AUG codons in ORF-1 are destroyed by point mutations, while partial enhancement is achieved by individual mutation of either ORF-1 AUG codon. These data suggest that FGF-5 synthesis requires the scanning of ribosomes past the two ORF-1 AUG codons. The introduction of these ORF-1 mutations into a eukaryotic FGF-5 expression vector increases its capacity to transform mouse NIH 3T3 cells up to 50-fold upon transfection. FGF-5 is secreted from transfected 3T3 cells and from human tumor cells as glycoproteins containing heterogeneous amounts of sialic acid. Glycosidase treatments suggest that the growth factor bears both N-linked and O-linked sugars.

Biosynthesis of human fibroblast growth factor-5

1991 ◽  
Vol 11 (4) ◽  
pp. 1840-1845
Author(s):  
B Bates ◽  
J Hardin ◽  
X Zhan ◽  
K Drickamer ◽  
M Goldfarb
Keyword(s):  
Growth Factor ◽  
Fibroblast Growth Factor ◽  
Human Fibroblast ◽  
Point Mutations ◽  
Human Tumor ◽  
Upstream Open Reading Frame ◽  
Fibroblast Growth ◽  
3T3 Cells ◽  
Reading Frame

We have analyzed the biosynthesis of human fibroblast growth factor-5 (FGF-5) at the translational and posttranslational levels. FGF-5 RNA synthesized in vitro can be translated in rabbit reticulocyte lysates to yield a 29,500-Da protein, which is consistent with the molecular weight predicted from the coding sequence. The efficiency of FGF-5 translation is dramatically enhanced if an upstream open reading frame (ORF-1) in the RNA is deleted or if both AUG codons in ORF-1 are destroyed by point mutations, while partial enhancement is achieved by individual mutation of either ORF-1 AUG codon. These data suggest that FGF-5 synthesis requires the scanning of ribosomes past the two ORF-1 AUG codons. The introduction of these ORF-1 mutations into a eukaryotic FGF-5 expression vector increases its capacity to transform mouse NIH 3T3 cells up to 50-fold upon transfection. FGF-5 is secreted from transfected 3T3 cells and from human tumor cells as glycoproteins containing heterogeneous amounts of sialic acid. Glycosidase treatments suggest that the growth factor bears both N-linked and O-linked sugars.

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