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Chinese Livestock and Poultry Breeding ›› 2026, Vol. 22 ›› Issue (5): 43-51.doi: 10.19543/j.cnki.1673-4556.20260407.001cstr: 32418.14.j.cnki.1673-4556.20260407.001

• Biotechnology • Previous Articles     Next Articles

Analysis of molecular characteristics of the CDS region of the sheep CYP17A1 gene and its expression in testes at different developmental stages

Yuxin Cai(), Meijie Wang, Xingwang Liu, Jialiang Zhang, Fang Li, Haotong Yang, Yuxin Yang, Xiaoyi Zhang, Man Bai()   

  1. College of Animal Science and Medicine, Shenyang Agricultural University, ShenYang, 110866, Liaoning
  • Received:2025-10-10 Online:2026-03-26 Published:2026-06-17
  • Contact: Man Bai E-mail:18847908342@163.com;doctorbaiman@163.com

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Abstract:

Objective This study was conducted to investigate the dynamic expression characteristics of CYP17A1 during testicular development in Small-tail han sheep, clarify its biological function and expression pattern, and provide a theoretical reference for further research. Method In this study, multiple bioinformatics tools were used to analyze the molecular characteristics of sheep CYP17A1 and construct a phylogenetic tree. Meanwhile, quantitative real‑time polymerase chain reaction (qRT‑PCR) and Western blot were performed to detect the expression of CYP17A1 in the testes of Small‑Tail Han sheep at 2, 6, and 12 months of age. Result The full length of the CDS region of the sheep CYP17A1 gene is 1576 bp, and it shows the closest genetic relationship with ruminants such as goats and cattle. The gene encodes 509 amino acids. The molecular formula of the CYP17A1 protein is C₂₆₀₅H₄₁₄₀N₆₉₆S₁₅, with a molecular weight of 57.39 kDa and a theoretical isoelectric point of 8.60. It is a slightly hydrophilic protein that contains phosphorylation sites but has no signal peptide. The secondary and tertiary structures of CYP17A1 are mainly composed of α-helix, β-sheet, extended strand, and random coil. Both the CYP17A1 gene and its protein were expressed in testicular tissue at all three developmental stages, and their expression levels showed a gradually decreasing trend with increasing age: the highest expression was observed at 2 months of age, followed by 6 months of age, and the lowest at 12 months of age. Conclusion Taken together, these findings demonstrate that CYP17A1 is involved in and exerts an essential regulatory effect on testicular development in Small-tail han sheep.

Key words: Small-tailed han sheep, Testicular tissue, CYP17A1, Molecular characterization

CLC Number: 

  • S826

Table 1

Online bioinformatics analysis software for sheep CYP17A1 protein and their website URLs"

软件Software网址Website URL功能Function
Expasyhttp://web.expasy.org/protparam/理化性质分析
SignalPhttps://services.healthtech.dtu.dk/services/SignalP-6.0/信号肽预测
TMHMMhttps://services.healthtech.dtu.dk/service.php TMHMM-2.0跨膜结构域预测
NetPhoshttps://services.healthtech.dtu.dk/services/NetPhos-3.1/磷酸化位点预测
DictyOGlychttps://services.healthtech.dtu.dk/services/DictyOGlyc-1.1/O-（α）-GlcNAc糖基化位点预测
NetNGlychttps://services.healthtech.dtu.dk/services/NetNGlyc-1.0/N-糖基化位点预测
Prabihttps://npsa-prabi.ibcp.fr/cgi-bin/npsa_automat.pl page=/NPSA/npsa_server.html二级结构预测
SWISS-MODELhttps://swissmodel.expasy.org/三级结构预测
PSORT Ⅱhttps://psort.hgc.jp/form2.html亚细胞定位

Table 2

Primer information"

基因名称

Gene names

引物序列（5′→3′）

Primer sequence

退火温度

Annealing temperature/℃

产物片段长度

Product fragment length/bp

CYP17A1F：CACCAGAGACTCCATCACTAACTTG; R：GCGAGCATGTGTCTGTTTGAAAGC60120
β-actinF：CCATCGGCAATGAGCGGTTCC; R：CGTGTTGGCGTAGAGGTCCTTG60146

Table 3

Similarity of the sheep CYP17A1 gene CDS region sequence with other species"

物种名

Species name

NCBI登录号

NCBI login number

相似性

Similarity/%

山羊Capra hircusNM_001314145.199.15
牛Bos taurusNM_174304.396.93
白尾鹿Odocolieus virginianusXM_020897894.296.27
猪Sus scrofaNM_214428.185.62
驴Equus asinusXM_014852085.382.94
马Equus caballusNM_001082523.182.75
猫Felis catusNM_001009371.282.75
犬Canis lupus familiarisXM_038440431.181.76
智人Homo sapiensNM_000102.480.26
兔Oryctolagus cuniculusXM_002718585.577.91
鼠Mus musculusNM_007809.372.09

Fig. 1

Phylogenetic tree of the CYP17A1 gene CDS region in sheep and other species"

Table 4

Amino acid composition of sheep CYP17A1 protein"

氨基酸Amino acid数量Quantity比例Proportion/%
丙氨酸 Ala（A）326.3
精氨酸 Arg（R）224.3
天冬酰胺 Asn（N）254.9
天冬氨酸 Asp（D）244.7
半胱氨酸 Cys（C）51.0
谷氨酰胺 Gln（Q）234.5
谷氨酸 Glu（E）305.9
甘氨酸 Gly（G）275.3
组氨酸 His（H）152.9
异亮氨酸 Ile（I）367.1
亮氨酸 Leu（L）7013.8
赖氨酸 Lys（K）367.1
蛋氨酸 Met（M）102.0
苯丙氨酸 Phe（F）275.3
脯氨酸 Pro（P）285.5
丝氨酸 Ser（S）356.9
苏氨酸 Thr（T）234.5
色氨酸 Trp（W）71.4
酪氨酸 Tyr（Y）71.4
缬氨酸 Val（V）275.3

Fig. 2

Bioinformatics analysis results of sheep CYP17A1 proteinNote: A is for transmembrane region prediction, B is for signal peptide prediction, C is for phosphorylation site prediction, D is for O-glycosylation modification site prediction, E is for N-glycosylation modification site prediction, F is for secondary structure prediction, G is for tertiary structure prediction, H is for interacting protein prediction."

Fig. 3

Relative expression levels of the CYP17A1 gene in testes of young sheep at different agesNote: Using β-actin as the internal control; * means significant difference (P < 0.05), ** means extremely significant difference (P < 0.01). The following picture is the same."

Fig. 4

Western blot grayscale images of CYP17A1 protein in testes of different ages"

Fig. 5

Relative expression level of CYP17A1 protein in the testes of small tail han sheep at different monthly ages"

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