In vitro analysis of expression vectors for DNA vaccination of horses: the effect of a Kozak sequence

نویسندگان

  • Guðbjörg Ólafsdóttir
  • Vilhjálmur Svansson
  • Sigurður Ingvarsson
  • Eliane Marti
  • Sigurbjörg Torsteinsdóttir
چکیده

One of the prerequisite for developing DNA vaccines for horses are vectors that are efficiently expressed in horse cells. We have analysed the ectopic expression of the human serum albumin gene in primary horse cells from different tissues. The vectors used are of pcDNA and pUC origin and include the cytomegalovirus (CMV) promoter. The pUC vectors contain CMV intron A whereas the pcDNA vectors do not. Insertion of intron A diminished the expression from the pcDNA vectors whereas insertion of a Kozak sequence upstream of the gene in two types of pUC vectors increased significantly the in vitro expression in primary horse cells derived from skin, lung, duodenum and kidney. We report for the first time the significance of full consensus Kozak sequences for protein expression in horse cells in vitro.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

In silico cloning and bioinformatics study of Brucella melitensis Omp31 antigen in different mammalian expression vectors

Brucella melitensis, as a pathogenic gram-negative intracellular bacterium, causes brucellosis in animals and humans. According to literature, the B. melitensis outer membrane protein 31 (Omp31) is considered as an important vaccine candidate against brucellosis. The aim of the current study was to compare three different expression constructs containing B. melitensis Omp31 antigen using bioinf...

متن کامل

Assessment of humoral immune response of a Cytomegalovirus DNA-vaccine candidate in BALB/c mice

Introduction: Glycoprotein B (gB) is the major antigen for induction of humoral responses against human cytomegalovirus (HCMV) making it an attractive candidate for immune prophylaxis. In the present study, the humoral immune response of BALB/c mice to a truncated HCMV gB protein fused with GFP was evaluated. Methods: The truncated gB coding sequence was synthesized and cloned in pEGFPN1 eukary...

متن کامل

Cloning and Expression of Immunogenic Regions of EMA-1 Gene of Theileria equi From Infected Horses

Diversity among the pathogenic strains of Theileria equi (T. equi), a major agent of equine piroplasmosis, can affect the appropriate detection of parasite and host immunization. Production of recombinant surface proteins from an infected horse in natural endemic area provides a reliable tool for immunodiagnosis of parasite. Regarding this, the present study was targeted toward the cloning, exp...

متن کامل

HIV-Derived Lentiviral Vectors: Current Progress toward Gene Therapy and DNA Vaccination

Lentiviral vectors are promising gene delivery tools capable of transducing a variety of dividing and non-dividing cells, including pluripotent stem cells which are refractory for transduction by murine retroviruses. Although there is a growing debate on the safety of lentiviral vectors for gene transfer, in particular for those derived from human immunodeficiency viruses, type one (HIV-1) and ...

متن کامل

CLONING AND EXPRESSION OF LEISHMANOLYSIN GENE FROM LEISHMANIA MAJOR IN PRIMATE CELL LINES

Leishmanolysin is a worldwide disease that is caused by different species of the genus Leishmania. Leishmanolysin, One of the genes expressed by Leishmania, appears to be an ideal candidate for genetic vaccination. In this study, a full length sequence, which encodes Leishmanolysin functionally critical regions (amino acids 100-579), was cloned from a Leishmania strain endemic to Iran. Analysis...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Acta Veterinaria Scandinavica

دوره 50  شماره 

صفحات  -

تاریخ انتشار 2008