3 research outputs found
Carborundum-dependent entrance of<i> Eco</i>RI restriction enzyme into plant cells and specific cleavage of genomic DNA
684-689In a basic research to determine the morpho-molecular interactions of plant tissues with EcoRI DNA restriction enzyme, it was demonstrated that this protein is capable of entering the sunflower and maize leaf cells using a plant tissue-abrading material and cleaving the genomic DNA at specific sites. This was inferred from the analysis of morphological patterns of EcoRI-treated leaf areas as well as using some molecular tests, including the cleavage pattern analysis of genomic DNA isolated from treated locations followed by ligation of cleaved fragments into EcoRI site of a DNA cloning vector system. The overall results indicated that the specific restriction of genomic DNA may happen following the entrance of EcoRI protein most likely into the nucleus of plant cells
<span style="font-size:11.0pt;font-family: "Times New Roman","serif";mso-fareast-font-family:"Times New Roman";mso-bidi-font-family: Mangal;mso-ansi-language:EN-GB;mso-fareast-language:EN-US;mso-bidi-language: HI" lang="EN-GB">Functional fusion expression of sunflower multicystatin in <i>E. coli</i> and its comparison with a single domain cystatin</span>
375-379<span style="font-size:11.0pt;font-family:
" times="" new="" roman","serif";mso-fareast-font-family:"times="" roman";mso-bidi-font-family:="" mangal;mso-ansi-language:en-gb;mso-fareast-language:en-us;mso-bidi-language:="" hi"="" lang="EN-GB">Identification of the molecular structure and novel biophysiological
functions of plant cystatins or phytocystatins is of great interest in the
field of molecular biology. The important
requirements for these are the efficient production, purification and
correctly folded forms of these proteins. We report here the cloning, easy
expression and characterization of a sunflower multicystatin (SMC) as a
functional fusion protein in E. coli. For the first time, the amplified
cystatin coding region was expressed as a part of maltose-binding fusion
protein using pMALc2X over-expression vector in TB1 strain of
E. coli without affecting the recombinant bacterial growth. In
comparison to the previously prepared recombinant SMC (rSMC), a high amount
(~44 mg/L of bacterial cell culture) of purified fused SMC (fSMC) was obtained
using single-step purification method. fSMC strongly inhibited papain activity in
vitro as compared to Celosia single-domain cystatin. Purified
fSMC may be used for basic biochemical, pharmacological or clinical studies
without the cleavage of its fusion parts.</span