A primer for the amplification of the mitochondrial 16s rRNA gene of Clamidae shellfish

Abstract

The invention provides an amplification primer of psammobiidae shellfish mitochondrion 16S rRNA genes. 72 psammobiidae 16S rRNA sequences are obtained by the amplification of a universal primer and are compared and analyzed by using CLUSTALW software; a plurality of pairs of primers are designed in a definite conserved region by using primer design software Premier Premier 5; PCR (polymerase chain reaction) test is carried out repeatedly on 230 psammobiidae individuals under a system of 0.01ml; and finally based on the detection of AGE (Agarose Gel Electrophoresis), an efficient amplification primer of psammobiidae shellfish mitochondrion 16S rRNA genes containing degenerate amino acid. The amplification primer is characterized by being ZG16Sa5'-TATCAAAAACATGGCCTCCTGA-3'ZG16Sb5'-AGCTCTGATCGCGTAARATTTYA-3'. According to the amplification primer, the target fragments of different psammobiidae species can be amplified effectively, wherein the amplification efficiency reaches up to 97%; and affinis species difficult to distinguish can be identified accurately, so that the requirements of studies on the genetic diversity of the psammobiidae are satisfied. Therefore, the amplification primer provided by the invention plays an important role in resource protection and utilization, the diversity assessment, the genetic breeding and other aspects of the psammobiidae.

Description

technical field [0001] The invention relates to a primer for amplifying the mitochondrial 16S rRNA [16S ribosomal RNA (16small-subunitribosomal RNA)] gene of the shellfish of the family Ziyun clam. Background technique [0002] Corbiculacea belongs to Bivalvia, Heterodonta, Veneroida, all marine products, widely distributed in coastal China, Japan, Philippines and Indian Ocean, usually buried in Intertidal zone to shallow sea fine sand or muddy seabed. Clamaceae is a class of bivalve molluscs with high economic value. Many species are delicious and rich in protein and vitamins. They can be eaten fresh or made into dry products. They have high edible value and important economic value. . For example, Chinese purple clams, double-lined purple clams, purple colored blood clams and pointed purple clams are all rare and precious sea treasures. There are many species and rich diversity of shellfish in this family, and the identification is generally based on morphological chara...

AI Technical Summary

Problems solved by technology

[0004] Although the 16S rRNA sequences of species of Clamidae are relatively conservative, the interspecies variability is high, which makes the amplification efficiency of 16S rRNA non-specific amplification primers low, and some species cannot be amplified at all, which hinders the use of molecular techniques

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