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  • Molekulsko kloniranje in cDNA sekvenca inhibitorjev cisteinskih proteinaz iz konjske morske vetrnice (Actinia equina L.) : diplomsko delo = Molecular cloning and cDNA sequence of cysteine proteinase inhibitors from seaanemone (Actinia equina L.) : graduation thesis
    Barlič, Ariana
    Several inhibitors of cysteine proteinases from sea anemone (Actinia equina L.) have been isolated. However their amounts were insuficient for furthetigations. So far only their N-terminal sequence ... have been determined, as well as three tryptic fragments. In order to determine their entire primarystructure, we have isolated positive clones from a lambda gt11 cDNA library made from the sea anemone mRNA, by immunoscreening. The positive cDNA clone was purified and amplified and the labmda DNA was isolated. The phage DNA was further cut using BamHI restriction endonuclease, and the following agarose gel electrophoresis displayed a band that corresponded to the positivecDNA of approximately 900 bp. The latter was then ligated into the pUC19 plasmids opened at the BamHI site, and the resulting recombinant plasmidDNA was used to transform competent Escherichia coli. DHSa cells. Afterisolation of the plasmid DNA we analysed the size of the inserted fragment which, again, tumed out to be 900 bp long. Using the Sanger's method of sequencing, the nucleotide sequence was determined. EI1 clone of 888 bp codes for a protein of 231 amino acid residues. Computer analysis of the aminoacid sequence (EI) helped us to predict its secondary structure, as well as to calculate its hydropathic index. According to the mostly hydrophobic character of the first 32 residue region located before the N-terminus of mature protein implies that this region corresponds to the signal peptide. We compared the N- and C-termini of the deduced mature protein form and found that both ends are highly similar in amino acid sequence, which led us to the assumption that, through the course of evolution, the gene encoding the ancestor of the EI, has probably been duplicated. (Abstract truncatrd. at 2000 caracter).
    Vrsta gradiva - diplomsko delo
    Založništvo in izdelava - Ljubljana : [A. Barlič], 1995
    Jezik - slovenski, angleški
    COBISS.SI-ID - 6943193

Knjižnica/institucija Kraj Akronim Za izposojo Druga zaloga
Nacionalni inštitut za biologijo in BF, Oddelek za biologijo, Ljubljana Ljubljana BFBNIB v čitalnico 1 izv.
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