Journal: Journal of Vaccines & Vaccination

Article Title: Immunization of Female Mice with a Plasmid DNA Vaccine Coding Eight Repeats of Gonadotrophin Releasing Hormone(Gnrh-I) and Eight T-Helper Epitopes Suppress Fertility In Vivo

doi: 10.4172/2157-7560.1000282

Figure Lengend Snippet: Figure 2: Western blotting detection of GnRH-I fusion protein obtained from transfected COS 1 cells and labelled with anti- GnRH-I (left) and anti V5 antibodies (right). The first [10], second [6] and the new vaccines were used to transfect COS 1 cells. Lane 1 shows the Kaleidoscope ladder, lanes 2, 3 and 4 containing transfected COS1 cell lysate of the first, second and new vaccines respectively. Lanes 6, 7 and 8 containing transfected COS1 cell culture supernatant of the first, second and new vaccine repectively. Lane 5 and 9 containing non-transfected COS1 cell lysate and culture supernatant respectively (negative control) and lane 10 lacking both anti-GnRH-I (left) and anti V5 antibodies (right) in the Western blotting detection of the fusion protein. The newly engineered vaccine produce fusion protein (35.216 KD) detected both in cell lysate (lane 4) and cell culture supernatant (lane 8). GnRH-I fusion protein (17.138 KD) of the first vaccine (lane 2) was detected in the cell lysate (lane 6). GnRH-I fusion protein (18.871 KD) of the second vaccine was detected both in cell lysate (lane 3) and cell culture supernatant (lane 7). GnRH-I fusion protein was not detected in any of the control lane 5, 9 and 10.

Article Snippet: A Kaleidoscope (Precision Plus ProteinTM BIO-RAD) protein marker was included in the starting wells and the separated protein bands in the gels were transferred to a nitrocellulose membrane (HybondTMECL membrane, Amersham Bioscience).

Techniques: Western Blot, Transfection, Vaccines, Cell Culture, Negative Control, Control