DNA uptake by embryo could be one of the causes for creation transgenic animals through SMGT

سال انتشار: 1393
نوع سند: مقاله کنفرانسی
زبان: انگلیسی
مشاهده: 959

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شناسه ملی سند علمی:

CIGS13_1140

تاریخ نمایه سازی: 7 بهمن 1393

چکیده مقاله:

The uses of live sperm as carrier of the transgene into oocytes were idea that was predicted to be able to make a great development in transgenesis. However, due to its very low reproducibility, heretofore this technique has not been confirmed in scientific society for transgenesis and discusses about it continues. The aim of this study was assessment of chickenembryo ability in DNA uptake as one of the causes of creation transgenic animals in sperm mediated gene transfer (socalledSMGT) and following injection of transgene in the vicinity of embryonic cells. Four treatments including: 2μg pDB2plasmid mixed with 3μl 0.5 M EDTA (PE); 2μg pDB2 plasmid Combined with Lipofectamine 2000 at 1 to 3 ratio (PL); 2μgpDB2 plasmid Combined with Lipofectamine 2000 at 1 to 3 ratio at presence of 3μl 0.5 M EDTA (PLE) and depc water asa control group (C) were considered. Injection was performed in the vicinity of embryo by opening a window with 10–15 mm at the sharp edge of egg shell in the stage X. In each group, 50 fertilized eggs were used. During the incubation period, tissue samples from different parts of the embryo collected and after examining embryos under the light green fluorescent,DNA extracted and PCR was performed for pDB2 transgene. Results obtained from light green fluorescent and PCR were positive for embryos that belonging to PLE group. In conclusion, transgene could be uptake by chicken embryo under physiological condition and incubation of only non-viral vector is insufficient for in vivo transgenesis of chicken embryo.

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نویسندگان

Hamid-Reza Amini

Department of Animal Science, University College of Agriculture and Natural Resources, University of Tehran, Karaj.

Abbas Pakdel

Department of Animal Science, University College of Agriculture and Natural Resources, University of Tehran, Karaj.

Hossein Moradi Shahr-Babak

Department of Animal Science, University College of Agriculture and Natural Resources, University of Tehran, Karaj.

Shahin Eghbalsaeid

Department of Animal Science, Khorasgan (Isfahan) Branch, Islamic Azad University, Isfahan, Iran.