| 1. |
- Jarvius, Jonas, et al.
(författare)
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Oligonucleotide ligation assay
- 2003
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Ingår i: Methods in Molecular Biology. - 1064-3745 .- 1940-6029. ; 212, s. 215-28
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Tidskriftsartikel (refereegranskat)
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| 2. |
- Johansson, Björn, et al.
(författare)
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Multiple Gene Expression by Chromosomal Integration and CRE-loxP-Mediated Marker Recycling in Saccharomyces cerevisiae
- 2004
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Ingår i: Recombinant Gene Expression Reviews and Protocols ( Methods in Molecular Biology ; 267 ). - New Jersey : Humana Press. - 1940-6029 .- 1064-3745. - 9781588292629 - 1592597742 ; 267
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Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- Multiple gene expression can be introduced in a yeast strain with using only two markers by means of the two new vectors described, the expression vector pB3 PGK and the CRE recombinase vector pCRE3. The pB3 PGK has a zeocin-selectable marker flanked by loxP sequences and an expression cassette consisting of the strong PGK1 promoter and the GCY1 terminator. The gene of interest (YFG1) is cloned between the promoter and terminator of pB3 PGK. The pB3 PGK-YFG1 is integrated into the genome by a single restriction cut within the YFG1 gene and integrated in the YFG1 locus. The strain is further transformed with the pCRE3 vector. The CRE recombinase expressed from this vector removes the zeocin marker and makes it possible to use the pB3 PGK vector over again in the same strain after curing of the pCRE3 vector. The 2µ-based pCRE3 carries the aureobasidin A, zeocin and URA3 markers. pCRE3 is easily cured by growth in nonselective medium without active counterselection. The screening for loss of the chromosomal zeocin marker, as well as curing of the pCRE3 vector, is done in one step, by scoring zeocin sensitivity. This can be done because the zeocin marker is present in both the pB3 PGK and pCRE3. The S. cerevisiae pentose phosphate pathway genes RK11, RPE1, TAL1, and TKL1 were cloned in pB3 PGK and integrated in the locus of the respective gene, resulting in simultaneous overexpression of the genes in the xylose-fermenting S. cerevisiae strain TMB3001.
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| 3. |
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| 4. |
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| 5. |
- Rådström, Peter, et al.
(författare)
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Pre-PCR processing of samples.
- 2003
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Ingår i: Methods in Molecular Biology. - 1064-3745 .- 1940-6029. ; 216
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Tidskriftsartikel (refereegranskat)
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| 6. |
- Degerman, Eva, et al.
(författare)
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Methods to study phosphorylation and activation of the hormone-sensitive adipocyte phosphodiesterase type 3B in rat adipocytes
- 2001
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Ingår i: Methods in Molecular Biology. - New Jersey : Humana Press. - 1940-6029. ; 155, s. 167-180
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Tidskriftsartikel (refereegranskat)abstract
- Cyclic nucleotide phosphodiesterases (PDEs) include a large group of structurally related enzymes that are responsible for the hydrolysis of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP). These enzymes belong to at least nine related gene families (PDEs 1–9) (1–5), which differ in their primary structures, affinities for cAMP and cGMP, responses to specific effectors, sensitivities to specific inhibitors, and regulatory mechanisms. The PDE3 family (6) consists of two subfamilies, PDE3A and PDE3B, which exhibit tissue-specific distribution; grossly, PDE3A enzymes are expressed in the cardiovascular system, and PDE3B enzymes in insulin-sensitive cells, such as hepatocytes (7) and adipocytes (6), and also in pancreatic β-cells (8). One characteristic of PDE3s involves their phosphorylation and activation in response to insulin, as well as to agents that increase cAMP in adipocytes (6), hepatocytes (7), and platelets (9–11), and in response to insulin-like growth factor-1 (IGF-)1 in pancreatic β-cells (8).
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| 7. |
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| 8. |
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| 9. |
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| 10. |
- Flodby, P, et al.
(författare)
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Knockout mice and steroid receptor research
- 2001
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Ingår i: Methods in molecular biology (Clifton, N.J.). - New Jersey : Humana Press. - 1064-3745. ; 176, s. 201-26
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Tidskriftsartikel (refereegranskat)
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