Nghiên cứu đặc điểm của gen chalcone isomerase phân lập từ cây sắn dây (Pueraria montana var. lobata)

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Tác giả: Thị Thu Huê Huỳnh, Tiến Dũng Nguyễn, Thị Bích Ngọc Trần

Ngôn ngữ: Vie

Ký hiệu phân loại: 633 Field and plantation crops

Thông tin xuất bản: Khoa học và công nghệ Việt Nam, 2021

Mô tả vật lý: 64 - 68

Bộ sưu tập: Metadata

ID: 414053

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Chalcone isomerase (CHI) được biết là enzyme quan trọng tham gia vào quá trình sinh tổng hợp các hoạt chất như flavonoid, isoflavonoid và anthocyanin. Enzyme này đã được nghiên cứu ở một số loài thực vật họ đậu, tuy nhiên ở Việt Nam chưa có nghiên cứu nào công bố về gen mã hóa CHI của cây sắn dây Pueraria montana var. lobata (P. lobata). Nghiên cứu này được thực hiện nhằm cung cấp thông tin, đặc điểm về gen mã hóa CHI thông qua việc nhân gen bằng RT-PCR, giải trình tự nucleotide, so sánh phân tích đặc điểm vùng chức năng trên trình tự gen và amino acid. Kết quả giải trình tự gen CHI bằng phương pháp Sanger cho thấy, vùng CDS có chiều dài 672 bp mã hóa cho 224 amino acid. So sánh trình tự nucleotide và amino acid giữa gen CHI của P. lobata và gen tham chiếu cùng loài (D63577.1) cho thấy mức độ tương đồng là 99,7%. 2 thay đổi nucleotide trên gen CHI làm thay đổi amino acid nhưng không nằm trong vị trí hoạt động bảo thủ của enzyme. Các vị trí hoạt động là một số amino acid bảo thủ (như liên quan đến liên kết hydro hoặc trung tâm liên kết cơ chất) cũng hiện diện trong gen CHI của P. lobata. Sử dụng SWISS-MODEL xây dựng mô hình cấu trúc protein cho thấy, protein CHI của P. lobata tương thích gần nhất với protein này của Medicago sativa - đã được xác định cấu trúc, với các vùng cấu trúc xoắn α và phiến gấp β hoàn toàn tương đồng. , Tóm tắt tiếng anh, Chalcone isomerase (CHI) is well-known as an important enzyme in the biosynthetic pathways such as flavonoid, isoflavonoid, and anthocyanin biosynthesis. The enzyme was investigated in some kinds of plants in Fabaceae but no research was conducted about the CHI gene of Pueraria montana var. lobata (P. lobata) in Vietnam. In order to provide more information and characterisation of the gene, our study isolated the CHI gene by RT-PCR and Sanger sequencing. The sequence of the CHI gene was analysed with nucleotide and deduced amino acid sequences to find the main domains. A full-length CDS of CHI gene from P. lobata is 672 bp encoded 224 amino acids. By using bioinformatic tools to compare, the isolated gene shared 99.7% homology with the same species reference (code D63577.1). Two different nucleotides in the gene were altered the amino acids in the protein, but the differences have not happened in active sites. Additionally, the conserved amino acids related to active catalysis of a hydrogen bond network also appeared in the P. lobata CHI gene. SWISS-MODEL was used to build the complete protein modeling showing that P. lobata CHI protein was the most similar with CHI of Medicago sativa - was defined structure in which all alpha-helix and beta-helix were completely homologies.

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