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| Cloning and Expression Analysis of SOX9L Gene in Pomacea canaliculata |
| ZHANG Shuang, SU Jing-Jing, WANG Xin-Hui, ZHANG Ying, LIU Feng-Yu, LIU Jing-Qi, YUAN Zi-Xia, LYU Jia-Li, SHANG Kai-Lin, YANG Shou-Bao* |
| School of Life Science, Shaoxing University, Shaoxing 312000, China |
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Abstract Pomacea canaliculata is one of the 16 most harmful invasive alien species in China. SRY-related HMG box gene 9 (SOX9) is a subgroup of SOXE genes associated with early embryonic development. In order to explore the function of the SOX9 gene of snails, the cDNA sequence of the SOX9 like gene from snail P. canaliculata (named as PcSOX9L, GenBank No. OP564918) was cloned by RT-PCR on the basis of transcriptome sequencing. Using P. canaliculata of different developmental stages (rankⅡ~ⅣP. canaliculata) as materials, the bioinformatics as well as expression characteristics of PcSOX9L were analyzed. The results showed that PcSOX9L gene contained a 1 701 bp open reading frame, encoding a hypothetical protein of 566 amino acids, and contained 2 conserved domains, namely Sox-N and HMG-box-domain; the amino acids of PcSOX9L and other animal SOX9 proteins showed that the sequence similarity ranged from 52.72% to 82.84%. qRT-PCR analysis showed that PcSOX9L gene was expressed in various tissues including gonad, liver, gill, mantle, and pleopod of rank Ⅳ P. canaliculata, as well as the albumin gland of female rank Ⅳ P. canaliculata. Among them, the expression level of PcSOX9L was the highest in the liver of male rank Ⅳ P. canaliculata, and was extremely significantly higher than the same tissues of female rank Ⅳ P. canaliculata (P<0.01). No significant difference was detected between the gonad of male and female rank Ⅳ P. canaliculata. The PcSOX9L was expressed in 3 different developmental stages, with the highest expression in rank Ⅱ P. canaliculata; at the same time, the changes of PcSOX9L expression in 3 different developmental stages toward male rank Ⅳ P. canaliculata in descending order as rank Ⅱ P. canaliculata>male rank Ⅳ P. canaliculata>>rank Ⅲ P. canaliculata; and the changes of PcSOX9L expression in 3 different developmental stages toward female rank Ⅳ P. canaliculata in descending order as rank Ⅱ P. canaliculata>rank Ⅲ P. canaliculata>female rank Ⅳ P. canaliculata, which indicated that this gene might play an important role in the early embryonic development and sex determination of P. canaliculata. The present results provide basic data for further analysis of the mechanism of SOX9 gene in the gonad development and sex determination of P. canaliculata.
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Received: 13 October 2022
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Corresponding Authors:
* shiouby@usx.edu.cn
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