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Structure and function of cyclophilins

100%

Nuc K., Slomski R.

Cyclophilins (CyPs) constitute a large class of highly conserved, ubiquitous PPIases (EC 5.2.1.8), which together with FK560 binding proteins (FKBP) and parvulins belong to a superfamily of immunophilins. These three classes of proteins are easily distinguishable by their selective interactions with immunosuppressive drugs. Cyclophilins are targets for cyclosporin A (CsA), parvulins bind juglone (5-hydroxy-1.4-naphthoquinone) and FKBP are inhibited either by FK560 drug or by rapamycin. Despite the lack of structural similarity all these proteins have been shown to act as peptidylprolyl cis-trans isomerases. In all cases binding of the drug inhibits their PPIase activity. Distinct isoforms of cyclophilins have been localized in the cytoplasm, nucleus, mitochondria, chloroplasts and endoplasmic reticulum.

Homology of DNA sequences encompassing the malignant hyperthermia mutation site in the human, porcine, and zebrine ryr1 gene

61%

Gronek P., Slomski R., Lisiecka D., Plawski A., Nuc K., Banasiewicz T.

Journal of Applied Genetics

1998

vol. 39

issue 3

275-279

The RYR1 gene encoding the Ca2+ channel of sarcoplasmic reticulum of human skeletal muscle has been cloned and its nucleotide sequence has been determined earlier. We have used the polymerase chain reaction single strand conformation polymorphism (PCR-SSCP), and sequencing analysis for human, porcine (Sus scrofa), and zebrine (Equus grevyi) ryanodine receptor (ryr1) gene. The fragment of exon 17 of the ryr1 gene was characterized by a high homology between all the analysed species (substitution of a nucleotide is underlined): porcine ryr1 1834GTG GCC GTG CGC TCC AAC CAA GAT CT1859 human RYR1 1831GTG GCC GTG CGC TCC AAC CAA GAT CT1856 zebrine ryr1 GTG GCC GTG CGC TCC AAC CAA GAC CT.

Expressive gene structures of proper and modified genes

42%

Lipinski D., Szalata M., Kalak R., Plawski A., Nuc K., Kala M., Juzwa W., Slomska K., Gronek P., Jura J., Smorag Z., Pienkowski M., Slomski R.

Biotechnologia

2003

issue 1

48-73

The genetic construct WAP 6xHisHGH containing the gene encoding human growth hormone (hGH) and WAP promoter expressed in mammary gland of animals was prepared. The 5? end of the gene was modified by the addition of sequence encoding six histidine residues and the sequence recognized by thrombin. In this way, the growth hormone can be easily purified by affinity chromatography and cleaved with thrombin to an active form. In the next step, the genetic construct was introduced by microinjection into male pronuclei of fertilized oocytes. Transgene was detected in male rabbit of F0 generation (number 61). Twelve offspring of founder rabbit of generation F1 indicated transgene sequences. The presence of growth hormone was revealed in the samples of milk accumulated during the lactation of females of F1 generation. The genetic constructs containing chain 1 and chain 2 of Feld1, and the major allergen produced by cat (Fedlis domesticus) were prepared. Both genes were inactivated by introduction into the sequences a positive selectable marker aminoglycoside phosphotransferase (resistant to neomycin). Outside the region of homology to Feld1 chain 1 and chain 2 genes, the negative selectable marker ? thymidine kinase gene was introduced. The genetic constructs pNTKFd1 and pNTKFd2 can be used in further experiments involving the inactivation of Feld1 genes in cat cells. Both genes were modified by site-directed mutagenesis using megastarter with Stop codon for premature termination of translation. The presence of mutation was confirmed by sequencing. The genetic constructs with human hGH gene and cat Feld1 gene were introduced into the bovine and cat fetal fibroblasts respectively in co-transfection with plasmid pGT-N29 containing positive selectable marker by lipofection, precipitation and electroinjection methods. After the selection, surviving cells were subjected to further molecular analysis. The stabile incorporation of the genetic constructs WAP 6xHisHGH and WAPHGH into the genome were observed.

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2 Biotechnologia

1 Journal of Applied Genetics

1 2003

1 2002

1 1998

3 Nuc K.

3 Slomski R.

2 Gronek P.

2 Plawski A.

1 Banasiewicz T.

1 Jura J.

1 Juzwa W.

1 Kala M.

1 Kalak R.

1 Lipinski D.

1 Lisiecka D.

1 Pienkowski M.

1 Slomska K.

1 Smorag Z.

1 Szalata M.

Search results

Structure and function of cyclophilins

Homology of DNA sequences encompassing the malignant hyperthermia mutation site in the human, porcine, and zebrine ryr1 gene

Expressive gene structures of proper and modified genes