A β-glucosidase with increased resistance to pepsin
A technology of glucosidase and pepsin, applied in the direction of enzymes, hydrolases, glycosylases, etc.
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0028] Embodiment 1: the synthesis of β-glucosidase gene
[0029] The present invention adopts the gene of wild-type β-glucosidase derived from Trichoderma viride (GenBank registration number is FJ882071.1), which is synthesized by Jinweizhi Company (other commercial companies with full gene synthesis can also complete it).
Embodiment 2
[0030] Embodiment 2: the connection of β-glucosidase gene (bgL1) and cloning vector Taox+PgHT+BBPB
[0031] 1. The pGH plasmid containing the bgL1 target gene synthesized by the whole gene and the cloning vector Taox+PgHT+BBPB were respectively digested with restriction endonucleases EcoRI and SpeI / XbaI at 37°C for 30 minutes, and the digestion conditions were as follows:
[0032]
[0033] 2. The digested products were subjected to 1% agarose gel electrophoresis and the two target fragments were respectively recovered, and T 4 DNA ligase connection, the connection system is as follows:
[0034]
[0035]
[0036] Use DNA ligase to ligate at 16°C for 16 hours, transform the ligation product into DH5α competent cells and amplify it, then use a plasmid extraction kit to extract the plasmid, digest it with EcoRI and PstI, and run electrophoresis results, showing two bands of 3.8kb and 7.0kb , indicating that the connection was successful, and determined to be the β-glucos...
Embodiment 3
[0037] Embodiment 3: gene fragment Paox+SS1 is connected with cloning vector M+Taox+PgHT+PB
[0038] 1. The gene fragment Paox+SS1 was transferred from the cloning vector Paox+SS1+PB preserved in this research institute, and obtained by double digestion with EcoRI and SpeI endonucleases, purification and recovery;
[0039] 2. The cloning vector M+Taox+PgHT+PB was obtained from Example 2, and the connection method between the gene fragment Paox+SS1 and the cloning vector M+Taox+PgHT+PB was the same as in Example 2.
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More