猪类无精症缺失基因DAZL的cDNA全长克隆、组织表达和亚细胞定位

doi:10.11843/j.issn.0366-6964.2021.03.012

Abstract

Abstract: The purpose of this study was to clone the full-length cDNA sequence of deleted in azoospermia like (DAZL) from Banna mini-pig inbred line (BMI), obtain its multi-tissue mRNA expression patterns, characterize the protein structure, and acquire the subcellular localization of DAZL. The tissue samples were collected from 10-month-old adult BMI boar and the full-length cDNA of DAZL gene was cloned from BMI using RACE and RT-PCR technologies. The multi-tissue mRNA transcription levels of DAZL were analyzed using qPCR. The structural characteristics and conserved domains of DAZL protein were analyzed using online tools. The subcellular localization of DAZL protein in the ST cell was detected by in vitro cell transfection technology. The results showed that the full-length cDNA sequence of DAZL was 2 985 bp (KU705632) with an 888 bp CDS encoding 295 amino acids (AOC89050). The BMI DAZL gene was located on chromosome 13 of BMI genome and contained 11 exons. Multi-tissue qPCR results showed that DAZL mRNA was exclusively and highly expressed in the testis. Further bioinformatics analysis showed that DAZL protein contained two conserved domains, RMP and DAZ, that were homologous with other mammals, with random coil structures more than 50%. Evolutionary analysis showed that the amino acid sequence of BMI DAZL was highly conserved and shared the closest genetic relationship with Bovidae. Fluorescence co-localization results of pEGFP-C1-DAZL transfected ST cells showed that DAZL protein was localized in the nucleus. The sequence characterization, mRNA expression, protein structure and location of BMI DAZL gene were clarified at DNA, mRNA and protein levels, respectively. This study will lay the foundation for further functional studies of DAZL during spermatogenesis in BMI.

Key words: Banna mini-pig inbred line (BMI), deleted in azoospermia like (DAZL), gene cloning, RACE, tissue expression, subcellular localization

CLC Number:

S828.3

WANG Pei, WANG Lina, HUO Hailong, ZHANG Xia, ZHAO Xiao, WANG Xuefei, HUO Jinlong. cDNA Full-length Cloning, Tissue Expression and Subcellular Location of DAZL Gene in Pig[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(3): 683-692.

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cDNA Full-length Cloning, Tissue Expression and Subcellular Location of DAZL Gene in Pig

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