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| Cloning of QKI Gene Family in Goat (Capra hircus) and Its Expression Pattern in Skeletal Muscle Development |
| XUE Ya-Nan, ZHAI Hong-Fan, TANG Min, YANG Chen-Yu, ZHOU Xin-Yi, LI Li, ZHANG Hong-Ping, ZHAN Si-Yuan* |
| Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu 611130, China |
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Abstract Quaking (QKI) is a member of RNA-binding protein family, and involved in the regulation of cell proliferation, differentiation and apoptosis. In order to explore the biological functions of QKI family genes in goat (Capra hircus) muscle development, Jianzhou big-eared goat was used as the research object in present study. The coding sequences of goat QKI family genes were obtained by TA cloning and analyzed by bioinformatics. qRT-PCR and Western blot were used to detect the expression of QKI family genes in different tissues (brain, heart, liver, spleen, lung, kidney, longissimus dorsi, semimembranosus, semitendinosus, gastrocnemius, psoas major, and adductor) of goats (3 d after birth) and developmental stages (embryonic 90 d, 105 d, 135 d; and 3 d, 150 d after birth) of the semimembranosus muscle, and their expression during the proliferation and differentiation of goat skeletal muscle satellite cells (SMSCs). The results showed that the complete coding sequences of goat QKI-5 (GenBank No. MW651981), QKI-6 (GenBank No. MW651982), and QKI-7 (GenBank No. MW651983) genes were obtained by amplification, with the length of 1 026, 978, and 960 bp, which encoding 341, 325, and 319 amino acids, respectively. Nucleotide sequence alignment revealed that QKI-5, QKI-6, and QKI-7 were highly conserved among species, and had the closest phylogenetic relationship with sheep (Ovis aries) and cattle (Bos taurus). The secondary and tertiary structures of goat QKI-5, QKI-6, and QKI-7 protein were mainly composed of α-helix and random coil, and there was a common conserved functional domain, KH (K homology) domain. The results of qRT-PCR showed that the expression of QKI-5, QKI-6, and QKI-7 genes in goat skeletal muscle tissues was significantly higher than that in other tissues (P<0.05); In the semimembranosus muscle of different developmental stages, QKI-5, QKI-6, and QKI-7 showed a trend of increasing first and then decreasing, with the highest expression at 3 d after birth (P<0.05). Western blot results showed that the expression of QKI-5 protein was only detected in goat skeletal muscle tissue, and mainly expressed after birth. Furthermore, QKI-5, QKI-6, and QKI-7 genes showed increasing expression tendency during the proliferation and differentiation of goat SMSCs, and reached the peak at 7 d after differentiation (P<0.05). The present study cloned and obtained the complete coding sequences of goat QKI-5, QKI-6, and QKI-7 genes, and revealed their high expression in goat skeletal muscle tissues and increasing expression patterns during proliferation and differentiation of goat SMSCs, indicating that QKI family genes might play an important role in goat skeletal muscle development. The present study provides basic data for further investigation of molecular mechanism on goat muscle development regulated by QKI.
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Received: 18 December 2020
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Corresponding Authors:
* siyuanzhan@sicau.edu.cn
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