کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
4371678 1302535 2008 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Toxoplasma gondii: Molecular cloning and characterization of a nitric oxide synthase-like protein
موضوعات مرتبط
علوم زیستی و بیوفناوری ایمنی شناسی و میکروب شناسی انگل شناسی
پیش نمایش صفحه اول مقاله
Toxoplasma gondii: Molecular cloning and characterization of a nitric oxide synthase-like protein
چکیده انگلیسی

Toxoplasma gondii has a nitrite production and a putative nitric oxide synthase (NOS) motif genomic sequence. In order to demonstrate that this sequence is functional and could be involved in the metabolism of l-arginine derivatives, we constructed a baculovirus carrying the previously identified Toxoplasma NOS-like DNA sequence. The recombinant protein was expressed into insect Sf9 cells and his activity was tested in serial microplate colorimetric assays. The protein produced 21 nmol/min/ml nitrites per microgram of protein and followed Michaelis–Menten kinetics, with a Km for l-arginine of 2.3 mM. Furthermore, the optimal pH, temperature and incubation time for the recombinant Toxoplasma NOS-like protein were established. Toxoplasma NOS runs as a band of 11.6 kDa on tricine–sodium dodecyl sulfate–polyacrylamide gel electrophoresis. Our results indicate that the recombinant protein derived from the putative genomic sequence, at the chromosome 1b of T. gondii, is able to produce nitrites from l-arginine as substrate.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Experimental Parasitology - Volume 119, Issue 3, July 2008, Pages 358–363
نویسندگان
, , , , ,