Metarhizium anisopliae protease pr1c and its gene and application
A technology of Metarhizium anisopliae and protease, applied in the field of agricultural biology, can solve the problems of not being able to prevent in advance, relying on chemical pesticides, etc., and achieve the effect of improving efficiency and increasing virulence
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Embodiment 1
[0031] Example 1 Clone Metarhizium anisopliae Pr1C gene
[0032] Metarhizium anisopliae total RNA was extracted with Separation reagents were used to extract the RNA of Metarhizium anisopliae tissue samples, the reverse transcription kit was used for reverse transcription, the Pr1C gene was cloned, the specific primers were shown in Table 1, and the Metarhizium anisopliae protease with nucleotide sequence as shown in SEQ ID No.2 was obtained Pr1C gene.
[0033] Table 1 The specific primers for cloning Pr1C gene
[0034]
[0035] Metarhizium anisopliae protease Pr1C gene was digested, and the digested product was inserted into pET-21b expression vector to obtain plasmid pET-Pr1C.
[0036] The ligation product was transformed into Escherichia coli DH5a strain, screened according to the marker of the recombinant vector (anti-Amp), single spot was picked, a small amount of plasmid was extracted by alkaline lysis, and preliminary identification was performed by double enzyme ...
Embodiment 2
[0037] Embodiment 2 Measuring the survival rate of locusts after feeding Metarhizium anisopliae PR1C protein
[0038] The 3rd instar nymphs of locusts in the same growth state were selected for the experiment, and 3 treatment groups and 2 control groups were set up. The treatment groups were: PR1C + Metarhizium anisopliae strain 202 treatment, PR1C treatment, Metarhizium anisopliae strain 202 treatment; the control group was: Induced bacteria, blank group. Each treatment group and control group had 5 replicates, and each replicate had 30 locusts.
[0039] Put the selected locusts into a constant temperature incubator and cultivate them at 28°C with a light-to-dark ratio of 16h:8h. After starvation treatment for 12h, they were fed with artificial feed, and fresh wheat seedlings were replaced after 24h. The number of deaths per day was recorded. The results of the survival rate of locusts are shown in the table 2 and figure 2 :
[0040] Table 2 Comparison table of locust sur...
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