Tạo dòng và biểu hiện gen mã hóa kháng nguyên P102 của Mycoplasma Hyopneumoniae trong E.Coli BL21 (DE3)

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Tác giả: Thị Hương Đặng, Thị Bích Lân Đinh, Hữu Rin Đồng, Quốc Việt Lê, Viết Quân Lê, Thị Thu Hiền Nguyễn, Thăng Long Phùng

Ngôn ngữ: vie

Ký hiệu phân loại:

Thông tin xuất bản: Tạp chí Khoa học kỹ thuật Thú y, 2020

Mô tả vật lý: tr.38

Bộ sưu tập: Metadata

ID: 331082

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In this study, we successfully cloned and expressed a fragment of gene encoding P102 antigen of Mycoplasma hyopneumoniae isolated from lung samples of pigs raised in Thua ThienHue province, Viet Nam. The fragment of gene encoding P102 antigen was cloned into plasmid pGEM®-T Easy and expressed with vector pET200/D-TOPO in Escherichia coli BL21 (DE3) cells. The studied results showed that the fragment length of gene encoding P102 antigen was 912 bp, encoding a polypeptide chain with 304 amino acid residues, and 100% similarity to the polypeptide chain of P102 antigen in GenBank (accession number AAZ44196.1). Results of SDS-PAGE electrophoresis showed that molecular weight of 6xHis-P102 recombinant protein was about 38 kDaIn this study, we successfully cloned and expressed a fragment of gene encoding P102 antigen of Mycoplasma hyopneumoniae isolated from lung samples of pigs raised in Thua ThienHue province, Viet Nam. The fragment of gene encoding P102 antigen was cloned into plasmid pGEM®-T Easy and expressed with vector pET200/D-TOPO in Escherichia coli BL21 (DE3) cells. The studied results showed that the fragment length of gene encoding P102 antigen was 912 bp, encoding a polypeptide chain with 304 amino acid residues, and 100% similarity to the polypeptide chain of P102 antigen in GenBank (accession number AAZ44196.1). Results of SDS-PAGE electrophoresis showed that molecular weight of 6xHis-P102 recombinant protein was about 38 kDaTrong nghiên cứu này, chúng tôi đã tạo dòng và biểu hiện thành công đoạn gen mã hóa kháng nguyên P102 của Mycoplasma hyopneumoniae phân lập từ mẫu phổi của lợn thu thập tại tỉnh Thừa Thiên-Huế, Việt Nam. Đoạn gen mã hóa cho kháng nguyên P102 được tạo dòng vào plasmid pGEM®-T Easy, sau đó được biểu hiện bởi vector pET200/D-TOPO trong tế bào Escherichia coli BL21 (DE3). Kết quả nghiên cứu cho thấy đoạn gen mã hóa kháng nguyên P102 có chiều dài 912 bp, mã hóa một chuỗi polypeptide dài 304 amino acid, và tương đồng 100% với trình tự chuỗi polypeptide của kháng nguyên P102 đã được công bố trên GenBank (mã số AAZ44196.1). Kết quả điện di trên SDS-PAGE cho thấy protein tái tổ hợp 6xHis-P102 có khối lượng phân tử khoảng 38 kDa.

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