طراحی سازه ژنی ترکیبی جهت تولید نوترکیب باکتریوسین لاتروسپورولین و آنالیز بیوانفورماتیکی پیش ساز پروتئینی با هدف بررسی نقش انتهای آمینی دمین 2SH

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه پاتوبیولوژی، دانشکده دامپزشکی، دانشگاه شیراز، شیراز، ایران

2 گروه زیست شناسی، دانشکده علوم، دانشگاه شیراز، شیراز، ایران

چکیده

باکتریوسین­ ها پپتیدهای تولید شده توسط باکتری ­ها می­ باشند که فعالیت ضدمیکروبی این پپتیدها بر علیه باکتری ­ها و قارچ­ ها شناخته شده است. به­  منظور تولید مقادیر بالای باکتریوسین با استفاده از فناوری DNA نوترکیب و نیاز به خنثی کردن اثر سمی پپتید و تولید فرم محلول پروتئین می ­توان از برخی پپتیدها که به ­عنوان چاپرون ­ها عمل می ­کنند، استفاده نمود. لاتروسپورولین یک باکتریوسین 49 اسیدآمینه ­ای می ­باشد که به ­طور طبیعی از باکتری 9-Brevibacillus laterosporus GI ترشح می­ شود. در مطالعه حاضر، برای اولین بار از انتهای آمینی دمین 2SH به ­عنوان قطعه پپتیدی کمکی جهت تولید نوترکیب باکتریوسین لاتروسپورولین استفاده شد. در این مطالعه سازه ژنی ترکیبی به­ صورت پیش­ ساز پروتئینی حاوی دنباله هیستیدینی، انتهای آمینی دمین 2SH (2N-SH)، جایگاه برش پروتئاز انتروکیناز و در نهایت پپتید لاتروسپورولین طراحی گردید و به ­کمک آنالیزهای بیوانفورماتیکی اثر دمین 2N-SH بر حلالیت و سطوح آب گریز پروتئین هدف مطالعه شد. نتایج نشان داد دمین 2N-SH با دارا بودن سطوح مناسب آب گریز می­ تواند به ­عنوان یک چاپرون مناسب، مانع از مجتمع شدن و به­ هم ریختگی ساختاری غیرقابل برگشت باکتریوسین لاتروسپورولین گردد. به ­علاوه آنالیزهای انجام شده نشان داد امکان بیان پروتئین هدف با تاخوردگی مناسب در شرایط آزمایشگاهی وجود دارد. مطالعه حاضر نشان داد دمین 2N-SH می ­تواند به ­عنوان یک گزینه مناسب جهت تولید نوترکیب انواع باکتریوسین ­ها در باکتریاشریشیا کلی عمل نماید.

کلیدواژه‌ها


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