A universal primer and method for efficiently and conveniently obtaining the mitochondrial genome of Psyllidoidea insects and its application

Abstract

The invention discloses a general primer and method for efficiently and conveniently obtaining the mitochondrial genome of Psyllidoidea insects and an application thereof. The universal primers include 6 pairs of primers: Psyllid-1, Psyllid-2, Psyllid-3, Psyllid-4, Psyllid-5 and Psyllid-6, the nucleotide sequences of the upstream and downstream primers are respectively as SEQ ID NO.1~12 shown. The present invention provides for the first time a universal primer for amplifying the mitochondrial genome of Psyllidoidae insects, which can be used to obtain the entire mitochondrial genome information of Psyllidoidae insects; The complete mitochondrial genome information of insects belonging to the superfamily Psyllididae can be obtained with the first-generation sequencing. The amplification efficiency is high, fast, easy to operate, and cheap. The requirements for operating instruments and analysis equipment are low, and it is easy to implement. The process is not disturbed by the genomes of insects themselves and other endophytes, and has great application prospects.

Description

technical field [0001] The invention relates to the technical field of molecular markers, in particular to a general primer and method for efficiently and conveniently obtaining the mitochondrial genome of Psyllidoidea and its application. Background technique [0002] Mitochondria are the main organelles for most eukaryotic cells to oxidize and provide energy. Its existence is related to cell survival and apoptosis. There is a set of mitochondrial genomes in this organelle, which is independent of nuclear chromosomes (Ridley, 2006). In recent years, the whole mitochondrial genome has played an important role in the study of insect evolution and population, mainly because of the relatively small genome, small number of genes, low recombination rate, high base mutation rate, heritability and high relative concentration. , so it is a good molecular genetic marker for studying insect molecular classification, phylogenetic evolution, population inheritance and variation (Boore, ...

AI Technical Summary

Problems solved by technology

[0006] The previously reported universal primers for psyllid mitochondria were mainly used to amplify some regions of the cox1 gene, and the information on the entire mitochondrial genome could not be obtained; however, the universal primers for animal mitochondrial genome sequencing designed by Simon et al. (2006) were based on 10 Designed by comparing the mitochondrial genome sequences of 18 insects, 2 arthropods, 2 vertebrates and 3 invertebrates

Although this set of primers has been widely used, due to the large variation in the structure of the mitochondrial genome of animals and the high number of base mutation sites, the versatility of this set of primers is not high, and there are many shortcomings in use. Not all primer pairs can be used in the entire The mitochondrial genome amplification has a very good effect. The applicant found in the early stage that this set of universal primers is not suitable for insect samples of the Psyllidoidea, and the amplification effect is poor, especially in genes with high variability. It works, but it can't be used for actual research

Images