Construction of recombinant yeast expressing EpEX as a suitable candidate in cancer diagnosis and therapy
Message:
Article Type:
Research/Original Article (دارای رتبه معتبر)
Abstract:
Introduction

Epithelial cell adhesion molecule (EpCAM) is a membrane glycoprotein that is overexpressed on the majority of tumor cells of epithelial origin and thereby can be used as a target of immunodetection and immunotherapy of cancer. So, it is important to produce this protein in its native form. Interestingly, during the last two decades, the yeast Pichia pastoris (P. pastoris) has become a popular host for the production of recombinant proteins because it combines the advantages of both mammalian and prokaryotic expression systems. In this study, the Pichia expressing EpCAM extracellular domain (EpEX) was constructed.

Materials and Methods

The codon optimized gene encoding EpEX protein was cloned in the XhoI and XbaI restriction sites of the pPICZαB vector. The constructed plasmid was integrated into GS115 strain by electroporation. Positive clones were evaluated by PCR using AOX1 primers.

Results

Sequencing as well as restriction enzyme analysis utilizing XhoI and XbaI (3506, 843 bp bands), as well as BamHI (3651, 698 bp bands) confirmed construction of recombinant EpEX pPICZαB. PCR based screening results of integrants showed two bands (2200 and 1345 bp), when AOX1 primer set was used.

Conclusion

These findings imply that the engineered strain was constructed. The constructed strain can be used in EpEX recombinant protein production for diagnostic and therapeutic purposes.

Language:
Persian
Published:
Pages:
735 to 742
magiran.com/p2038983  
دانلود و مطالعه متن این مقاله با یکی از روشهای زیر امکان پذیر است:
اشتراک شخصی
با عضویت و پرداخت آنلاین حق اشتراک یک‌ساله به مبلغ 990,000ريال می‌توانید 70 عنوان مطلب دانلود کنید!
اشتراک سازمانی
به کتابخانه دانشگاه یا محل کار خود پیشنهاد کنید تا اشتراک سازمانی این پایگاه را برای دسترسی نامحدود همه کاربران به متن مطالب تهیه نمایند!
توجه!
  • حق عضویت دریافتی صرف حمایت از نشریات عضو و نگهداری، تکمیل و توسعه مگیران می‌شود.
  • پرداخت حق اشتراک و دانلود مقالات اجازه بازنشر آن در سایر رسانه‌های چاپی و دیجیتال را به کاربر نمی‌دهد.
دسترسی سراسری کاربران دانشگاه پیام نور!
اعضای هیئت علمی و دانشجویان دانشگاه پیام نور در سراسر کشور، در صورت ثبت نام با ایمیل دانشگاهی، تا پایان فروردین ماه 1403 به مقالات سایت دسترسی خواهند داشت!
In order to view content subscription is required

Personal subscription
Subscribe magiran.com for 50 € euros via PayPal and download 70 articles during a year.
Organization subscription
Please contact us to subscribe your university or library for unlimited access!