hmg1 Gene and Its Application in Molecular Detection of Microsporidium Bombyx mori

Abstract

The invention discloses a silkworm microsporidia HMG1 gene as well as a specific primer set used for rapidly detecting silkworm microsporidia and application thereof. The primer set comprises an upstream primer HMG1-s and a downstream primer HMG1-sR, wherein a nucleotide sequence of the upstream primer is shown in SEQ ID No.5, and a nucleotide sequence of the lower primer is shown in SEQ ID NO.6. A target gene of the detection primer is a high mobility protein gene HMG1 related to sex of the silkworm microsporidia, when the high mobility protein gene HMG1 is taken as the target gene for a silkworm tissue, especially silkworm egg microsporidia molecular detection, the characteristics of reliable detection result, easy operation, strong specificity and high sensitivity can be realized, and the high mobility protein gene HMG1 can be applied to high-sensitivity and rapid silkworm microsporidia molecular detection, especially early detection of the silkworm microsporidia, and has great significance in practical application.

Description

technical field [0001] The invention belongs to the technical field of insect pathogenic microorganism molecular detection. More specifically, it relates to HMG1 gene and its application in the molecular detection of silkworm Microsporidia. Background technique [0002] The study of silkworm microparticle disease (also known as silkworm microsporidiosis) began with a nationwide epidemic of microparticle disease in France in 1845. Pasteur determined that the "microparticles" he observed were the causative factor for microparticle disease, which Balbiani later identified as Bombyx mori Nosema bombycis (J.V. Maddox et al., 2000). Silkworm Microsporidia has two modes of infection: horizontal transmission and vertical transmission, among which vertical transmission causes great harm to the production of silkworm seeds in sericulture production, and at the same time has a great negative impact on the yield and quality of silk cocoons for breeding silkworms. Influence the develop...

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Problems solved by technology

Most of the target genes of the primers designed in the research of silkworm microsporidian PCR detection technology are also SSUrRNA, and the primers designed for other microsporidian genes are less or have poor sensitivity, so they are rarely reported

Baker et al (1995) and Terry et al (1999) designed PCR primer V1f / 530r based on the highly conserved region of SSU rRNA of similar species of Microsporidia, which can identify DNA templates of various species of Microsporidia and amplify about 450bp Specific target bands, but not specific to Bombyx mori Microsporidia, and the inventors found that using this primer to detect silkworm egg Microsporidia, the detection sensitivity is extremely low, suggesting that there may be a certain Inhibitory factors interfere with PCR amplification of DNA from No.

On the other hand, microsporidia parasitize in silkworm eggs, and the content of silkworm eggs is significantly higher than that of the microsporidia to be detected. In the extracted DNA samples, both DNAs exist at the same time, and the DNA of silkworm silkworm eggs seriously affects the detection. Interference, therefore, if you want to directly use silkworm egg DNA as a template for the detection of microsporidia, higher requirements are put forward for the detection

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