Mối quan hệ giữa mức độ biểu hiện gen PMI (manA) và áp lực chọn lọc tối ưu trong chuyển gen lúa Indica

Plant Cell Reports - Tập 33 - Trang 1081-1090 - 2014
Huaping Gui1, Xia Li1, Yubo Liu1, Kai Han1, Xianggan Li1,2
1Syngenta Biotechnology China Co., Ltd, Beijing, People’s Republic of China
2Syngenta Biotechnology Inc, Durham, USA

Tóm tắt

Một hệ thống lựa chọn mannose hiệu quả đã được thiết lập cho việc chuyển gen giống lúa Indica IR58025B. Các áp lực lựa chọn khác nhau đã được yêu cầu để đạt được tần suất chuyển đổi tối ưu cho các băng cassette đánh dấu PMI khác nhau. Nghiên cứu này được thực hiện nhằm thiết lập một hệ thống chuyển gen hiệu quả cho lúa Indica, giống IR58025B. Bốn tổ hợp của hai promoter, Actin 1 của lúa và Ubiquitin 1 của ngô, cùng với hai gen manA, gen tự nhiên từ E. coli (PMI-01) và gen tổng hợp được tối ưu hóa mã từ ngô (PMI-09), đã được so sánh dưới các nồng độ mannose khác nhau. Các áp lực lựa chọn khác nhau đã được yêu cầu cho các băng cassette gen khác nhau để đạt được tần suất chuyển đổi tối ưu tương ứng (TF). Tần suất chuyển đổi cao hơn như 54 và 53% đã được thu được khi sử dụng 5 g/L mannose cho việc chọn băng cassette prActin-PMI-01 và 7.5 g/L mannose cho việc chọn băng cassette prActin-PMI-09, tương ứng. Tần suất chuyển đổi là 67 và 56% đã được thu được khi sử dụng 7.5 và 15 g/L mannose cho việc chọn băng cassette prUbi-PMI-01 và prUbi-PMI-09, tương ứng. Chúng tôi kết luận rằng tần suất chuyển đổi cao hơn có thể đạt được cho các băng cassette gen khác nhau khi áp dụng áp lực lựa chọn tối ưu. Bằng cách điều tra mức độ biểu hiện PMI ở các callus và lá chuyển gen, chúng tôi thấy có một mối tương quan tích cực đáng kể giữa mức độ biểu hiện protein và áp lực lựa chọn tối ưu. Áp lực lựa chọn tối ưu cao hơn là cần thiết cho những cấu trúc mà tạo ra mức biểu hiện protein PMI cao hơn. Tỷ lệ bản sao đơn của những sự kiện chuyển gen cho băng cassette prActin-PMI-01 thấp hơn so với ba băng cassette còn lại. Chúng tôi suy đoán rằng một số sự kiện bản sao thấp với mức độ biểu hiện protein thấp có thể không sống sót được trong quá trình lựa chọn mannose.

Từ khóa

#chuyển gen lúa Indica #băng cassette gen PMI #áp lực lựa chọn #biểu hiện protein #tần suất chuyển đổi

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