Đánh giá sự biểu hiện của gen AtZAT12 trên cây Arabidopsis chuyển gen

0 Người đánh giá. Xếp hạng trung bình 0

Tác giả: Thị Tuyết Châm Lê, Anh Tuấn Trần, Ngọc Thắng Vũ, Thị Thúy Hằng Vũ

Ngôn ngữ: Vie

Ký hiệu phân loại: 576.5 Genetics

Thông tin xuất bản: Khoa học nông nghiệp Việt Nam 2020

Mô tả vật lý: 545-552

Bộ sưu tập: Metadata

ID: 406960

Thêm vào giỏ

ZAT12 là một yếu tố phiên mã đáp ứng stress có sự biểu hiện tăng cao khi cây chịu một số stress như oxy hóa, lạnh và nhiệt... ZAT12 liên kết đáp ứng thiếu sắt với stress oxy hóa thông qua tương tác trực tiếp và điều hòa âm tính với yếu tố phiên mã FIT. Trong nghiên cứu này, các dòng chuyển gen ZAT12 được điều khiển bởi promoter của chính gen ZAT12. Mục tiêu của nghiên cứu là tạo được các dòng chuyển gen AtZAT12 biểu hiện cao (ZAT12 Ox) đáp ứng lại với stress oxy hóa liên quan đến hấp thụ Fe. Chúng tôi tiến hành chọn lọc các dòng ZAT12 Ox bằng phương pháp quan sát hạt phát huỳnh quang, đọc trình tự và PCR. Các dòng đồng hợp tử được đánh giá sinh trưởng, sự biểu hiện ở mức phiên mã và dịch mã. Kết quả chọn được 4 dòng dương tính T0 (kí hiệu CH1-1, 1-4, 1- 10 và 1-14) có sinh trưởng khác biệt, trong đó sinh trưởng của dòng CH1-1 bị ức chế. Trong điều kiện thiếu Fe, các dòng ZAT12 Ox này có biểu hiện tăng cao, nhưng không tăng thêm khi xử lý thêm với H2O2. Sự biểu hiện gen ở mức độ dịch mã cũng đã được quan sát thấy ở dòng CH1-4 bằng Western Blot., Tóm tắt tiếng anh, ZAT12 is one of the transcription factors response to abiotic stress, which has had higher expression of ZAT12 under oxidative, cold, and heat stress etc. ZAT12 links iron deficiency and oxidative stress responses through direct interaction with and negative regulation of FIT. However, in this study, ZAT12 was regulated by ZAT12 native promoter. This study aimed to generate ZAT12 with overexpression (ZAT12 Ox) in response to oxidative stress and Fe uptake. The attempts of this study were further selected the transgene lines containing 2x35S::ZAT12 by observing the dry seed under fluorescent microscope gene sequencing and PCR. The growth and gene expression as well as ZAT12 protein regulation of 2x35S::ZAT12 lines (ZAT12 Ox) were evaluated. As the results showed, the 4 positive lines (CH1-1, 1-4, 1-10 and 1-14 lines) with different growth were generated, of which the growth of CH1-1 line was inhibited. Expression of AtZAT12 gene induced to response to Fe deficiency, but not to H2O2 supplemented treatment. ZAT12 protein of CH1-4 line also detected by Western Blot.

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