The invention discloses a method for constructing a
target protein interaction network based on high-
throughput sequencing. According to the method, a BD-bait and a prey
library are co-transformed into a Y2H
yeast strain, then culturing in a YPDplus culture medium at 30 DEG C for 4 hours, discarding the supernatant, adding 1mL of
sterile water medium for suspension, taking 100muL of suspension bacterial liquid and adding into 10ml of SD / -Leu / -Trp / -His / -Ade
liquid culture medium for shaking culture, sampling respectively after shaking culture for 24 hours, 72 hours and 80 hours, the sampled bacterial liquids are used as templates for PCR (
Polymerase Chain Reaction) amplification, carrying out
agarose gel electrophoresis to test the screening degree of the bacterial liquids, recycling products, carrying out high-
throughput sequencing, carrying out
gene abundance and
difference analysis after de novo
assembly, and taking a difference
gene as a
protein coding
gene interacting with a targetgene in the
library, so as to preliminarily construct an
interaction network of the
target protein. The method is also suitable for constructing a
target protein interaction network of species without
reference genes. The method simplifies the steps of a
yeast two-
hybrid screening
library, and the method is not limited by the number of clones screened once, and can find all interaction proteins of a target
protein in a single library.