A method for detecting the expression of the odorant-binding protein gene obp6 in Apis mellifera using fluorescent RT-PCR

Abstract

The invention discloses a method for detecting expression of an odorant-binding protein gene OBP6 of Italian bees by using fluorescent RT-PCR technology, and particularly relates to the technical field of biology. The method for detecting the expression of the odorant-binding protein gene OBP6 of the Italian bees by using the fluorescent RT-PCR technology comprises the following steps that (1), OBP6 genes are detected by using primers; (2), total RNA extraction is performed; (3), cDNA synthesis is performed; (4), real-time fluorescent PCR is performed; and (5), the relative expression quantityof OBP6 of the Italian bees is calculated before and after benomyl treatment. The real-time fluorescent RT-PCR adopted by the method for detecting the expression of the odorant-binding protein gene OBP6 of the Italian bees by using the fluorescent RT-PCR technology is compared with conventional PCR, and the real-time fluorescent PCR technology collects fluorescent signals by automated instruments, the subjectivity of visual judgment is avoided, and the sensitivity is further improved; and at the same time, the real-time fluorescent RT-PCR technology reacts in a fully closed mode without post-treatment of PCR, pollution is avoided, and the reliability and repeatability of the results are ensured. At the same time, the following tedious steps such as electrophoresis and quantitative scanning in routine PCR are eliminated, and the experiment time is greatly shortened.

Description

Technical field: [0001] The invention relates to the field of biotechnology, and relates to specific primers for detecting the expression of the Apis mellifera odorant-binding protein gene obp6, more specifically a method for detecting the expression of the Apis mellifera odorant-binding protein gene OBP6 by using a fluorescent RT-PCR technique. Background technique: [0002] In the long-term evolution process of insects, the environment they face is highly heterogeneous. The adaptation of insects to the heterogeneous environment is an important driving force for their survival and evolution. In order to survive and reproduce normally, insects must interact with the complex surrounding environment. produced good adaptability. Usually, the information relationship between individual insects and between insects and the environment is mainly regulated by chemical communication. The insect olfactory system is a highly specific and extremely sensitive chemical detector, which can...

AI Technical Summary

Problems solved by technology

Although it is not possible to fully elucidate how honeybee odorant-binding proteins (OBPs) participate in the olfactory perception mechanism of bees, the physiological and biochemical characteristics and binding characteristics of honeybee odorant-binding proteins have been clarified, which is helpful for grasping the olfactory perception mechanism of bees and explaining how honeybees feel and smell. Identifying odorous substances in the environment and their corresponding behavioral responses have certain guiding significance

[0005] In recent years, due to the excessive use of pesticides in agricultural and forestry production, the widespread use of antibiotics in the process of bee disease control, environmental pollution, interference from human activities, and destruction of vegetation, the ecological environment has been destroyed, and the immune system and olfactory system of bees have been destroyed. , so that bees are frequently attacked by diseases and parasites in the process of breeding, and even in European and American countries, bees have colony collapse syndrome, which has caused huge economic losses to beekeepers

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