种猪肺炎支原体的融合基因及其制备方法

• Format: Patent
• Language: Chinese
• Published: 02.10.2018
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

本发明公开了种猪肺炎支原体的融合基因及其制备方法，属于基因工程技术领域，本发明中所使用的P46和P65基因，直接根据NCBI GeneBank中发布的编码区序列，设计含有相应酶切位点的特异性引物。P46基因的PCR产物和原核表达载体pET-28a双酶切并连接，构建重组表达载体pET-28a-P46；再令重组表达载体pET-28a-P46和P65基因的PCR产物双酶切并连接，直接构建重组表达载体pET-28a-P46-P65。优势是不需要使用Linker连接P46和P65基因，直接经过双酶切连接就可得到重组表达载体pET-28a-P46-P65。重组载体构建过程相较于，使用Linker连接基因再与原核表达载体连接，过程简单并且易于得到，耗费时间明显减少。 The invention discloses a fusion gene of mycoplasma hyopneumoniae and a preparation method of the fusion gene, and belongs to the field of genetic engineering technology. P46 and P65 genes adopted in the invention can directly design specific primers containing corresponding restriction sites according to a coding region sequence released in NCBI GeneBank. A PCR product of the P46 gene is subject to double enzyme digestion and connection with a prokaryotic expression vector pET-28a, so as to construct a recombinant expression vector pET-28a-P46; and the recombinant expression vector pET-28a-P46 is subject to double enzyme digestion and connection with a PCR product of the P65 gene, so