绵羊CYP17A1基因CDS区分子特征分析及其在不同时期睾丸中的表达研究

doi:10.19543/j.cnki.1673-4556.20260407.001

摘要/Abstract

摘要：

目的为探究CYP17A1在小尾寒羊睾丸发育中的动态表达特征，明确其功能和表达规律，为后续深入研究提供参考。方法该研究分别通过不同的生物信息学软件对绵羊CYP17A1的分子特征展开研究，并构建系统发育进化树；同时，运用实时荧光定量PCR（qRT-PCR）与蛋白免疫印迹（Western Blot）技术检测CYP17A1在不同月龄（2、6、12月龄）小尾寒羊睾丸中的表达情况。结果绵羊CYP17A1基因CDS区全长1576 bp，与山羊和牛等反刍动物亲缘关系最近，共编码509个氨基酸，CYP17A1蛋白分子式为C₂₆₀₅H₄₁₄₀N₆₉₆S₁₅，分子量为57.39 kDa，理论等电点8.60，是一种具有磷酸化位点但不存在信号肽的轻微亲水性蛋白。二级结构和三级结构均以α-螺旋、β-转角、延伸链和无规卷曲构成；CYP17A1基因和蛋白在3个时期睾丸组织中均有表达，且表达量均随着月龄增长呈逐级下降的趋势，即2月龄表达量最高，6月龄的表达量次之，12月龄的表达量最低。结论由此可见，CYP17A1基因影响小尾寒羊睾丸发育，并且对睾丸发育起到重要的调控作用。

关键词: 小尾寒羊, 睾丸组织, CYP17A1, 分子特征分析

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

中图分类号:

S826

蔡雨欣, 王美桀, 刘兴旺, 张嘉良, 李放, 杨浩彤, 杨宇欣, 张笑怡, 白曼. 绵羊CYP17A1基因CDS区分子特征分析及其在不同时期睾丸中的表达研究[J]. 中国畜禽种业, 2026, 22(5): 43-51.

Yuxin Cai, Meijie Wang, Xingwang Liu, Jialiang Zhang, Fang Li, Haotong Yang, Yuxin Yang, Xiaoyi Zhang, Man Bai. Analysis of molecular characteristics of the CDS region of the sheep CYP17A1 gene and its expression in testes at different developmental stages[J]. Chinese Livestock and Poultry Breeding, 2026, 22(5): 43-51.

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参考文献 34

[1]	严桂芹. 种公羊的饲养繁育技术[J]. 养殖与饲料, 2024, 23(4): 61-63.
	YAN G Q. Breeding techniques of breeding rams[J]. Animal Breeding and Feed, 2024, 23(4): 61-63.
[2]	COX J F, JERIA E, BOCIC A, et al. Characterization of the productive performance of Highlander sheep in Southern Chile. II. Male reproductive traits[J]. Small Ruminant Research, 2015, 130: 189-192.
[3]	缑帅帅, 刘玲玲, 曹行, 等. 基于全基因组数据对新疆5个地方绵羊品种特异性SNP位点的筛选及鉴定[J]. 中国畜牧兽医, 2025, 52(10): 4754-4764.
	GOU S S, LIU L L, CAO H, et al. Screening and identification of specific SNPs for 5 local sheep breeds in Xinjiang based on whole-genome data[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(10): 4754-4764.
[4]	YANG J, WANG D F, HUANG J H, et al. Structural variant landscapes reveal convergent signatures of evolution in sheep and goats[J]. Genome Biology, 2024, 25(1): 148.
[5]	LUKIC B, CURIK I, DRZAIC I, et al. Genomic signatures of selection, local adaptation and production type characterisation of East Adriatic sheep breeds[J]. Journal of Animal Science and Biotechnology, 2023, 14(1): 142.
[6]	石东民, 肖海英. 提高小尾寒羊能繁母羊生产性能的要点[J]. 中国畜牧业, 2025(18): 44-45.
	SHI D M, XIAO H Y. Key points for improving reproductive performance of fertile ewes of small-tailed Han sheep[J]. China Animal Industry, 2025(18): 44-45.
[7]	张海涛. 不同营养水平对超早期断奶小尾寒羊母羊繁殖性能(发情时间)影响的研究[D]. 兰州: 甘肃农业大学, 2004.
	ZHANG H T. The study of effect of over-early weaning of small tail Han ewes reproductive performance (Oestrus time) at different nutrition levels[D]. Lanzhou: Gansu Agricultural University, 2004.
[8]	张亚男. 小尾寒羊母羊繁殖周期血液指标变化规律及其与FecB基因多胎性关联分析[D]. 泰安: 山东农业大学, 2023.
	ZHANG Y N. Changes of blood indexes in the reproductive cycle of small-tailed Han sheep ewes and their association with FecB gene prolificacy[D]. Taian: Shandong Agricultural University, 2023.
[9]	赵晏. 提高小尾寒羊能繁母羊繁殖率的养殖要点[J]. 养殖与饲料, 2024, 23(11): 57-60.
	ZHAO Y. Key points of breeding to improve the reproductive rate of small-tailed Han sheep[J]. Animal Breeding and Feed, 2024, 23(11): 57-60.
[10]	LI L, LIN W Q, WANG Z Y, et al. Hormone regulation in testicular development and function[J]. International Journal of Molecular Sciences, 2024, 25(11): 5805.
[11]	TANG W, ZHANG Y, WANG Z, et al. Key role of CYP17A1 in Leydig cell function and testicular development in Qianbei Ma goats[J]. Genomics, 2025, 117(1): 110937.
[12]	曾起, 胡蓓娟, 邱惠敏, 等. CYP17A1在池蝶蚌性腺发育中的调控作用[C]. //中国动物学会·中国海洋湖沼学会贝类学分会换届大会暨第二十一次全国贝类学术讨论会论文集. 昆明, 2024: 82.
	ZENG Q, HU B J, QIU H H, et al. Regulatory role of CYP 17A1 in gonadal development of Hyriopsis schlegelii[C]. //Abstracts of the General Election Congress of Malacology Branch of Chinese Zoological Society & Chinese Society for Oceanology and Limnology and the 21st National Malacology Academic Symposium. Kunming, 2024: 82.
[13]	AUCHUS R J. The backdoor pathway to dihydrotestosterone[J]. Trends in Endocrinology & Metabolism, 2004, 15(9): 432-438.
[14]	张远, 陈祥, 张艳, 等. CYP17A1基因敲低对山羊睾丸间质细胞睾酮合成和睾丸发育相关基因表达的影响[J]. 农业生物技术学报, 2023, 31(12): 2535-2544.
	ZHANG Y, CHEN X, ZHANG Y, et al. Effect of CYP17A1 gene knockdown on expression of genes related to testosterone synthesis and testicular development in goat(Capra hircus) leydig cells[J]. Journal of Agricultural Biotechnology, 2023, 31(12): 2535-2544.
[15]	陈思. 不同FecB基因型小尾寒羊下丘脑和垂体转录组学研究[D]. 北京: 中国农业科学院, 2022.
	CHEN S. Transcriptome Analysis of small tail han sheep hypothalamus and pituitary gland with different FecB genotypes[D]. Beijing: Chinese Academy of Agricultural Sciences, 2022.
[16]	BURRIS-HIDAY S D, SCOTT E E. Steroidogenic cytochrome P450 17A1 structure and function[J]. Molecular and Cellular Endocrinology, 2021, 528: 111261.
[17]	YADAV R, PETRUNAK E M, ESTRADA D F, et al. Structural insights into the function of steroidogenic cytochrome P450 17A1[J]. Molecular and Cellular Endocrinology, 2017, 441: 68-75.
[18]	PAYNE A H, HALES D B. Overview of steroidogenic enzymes in the pathway from cholesterol to active steroid hormones[J]. Endocrine Reviews, 2004, 25(6): 947-970.
[19]	CUI Y L, ZHENG Q C, ZHANG J L, et al. Molecular dynamic investigations of the mutational effects on structural characteristics and tunnel geometry in CYP17A1[J]. Journal of Chemical Information and Modeling, 2013, 53(12): 3308-3317.
[20]	MILLER W L, AUCHUS R J. The molecular biology, biochemistry, and physiology of human steroidogenesis and its disorders[J]. Endocrine Reviews, 2011, 32(1): 81-151.
[21]	ZHANG L H, RODRIGUEZ H, OHNO S, et al. Serine phosphorylation of human P450c17 increases 17, 20-lyase activity: implications for adrenarche and the polycystic ovary syndrome[J]. Proceedings of the National Academy of Sciences of the United States of America, 1995, 92(23): 10619-10623.
[22]	AUCHUS R J, LEE T C, MILLER W L. Cytochrome b 5 augments the 17, 20-lyase activity of human P450c17 without direct electron transfer[J]. Journal of Biological Chemistry, 1998, 273(6): 3158-3165.
[23]	LI Y, ZHANG J, ZHANG X, et al. Expression patterns of steroidogenic enzymes in the developing testes of Holstein bulls[J]. Animal Reproduction Science, 2020, 220: 106353.
[24]	O’SHAUGHNESSY P J, BAKER P J, MONTEIRO A, et al. Developmental changes in human fetal testicular cell numbers and messenger ribonucleic acid levels during the second trimester[J]. The Journal of Clinical Endocrinology and Metabolism, 2007, 92(12): 4792-4801.
[25]	PLANT T M. Hypothalamic control of the pituitary-gonadal axis in higher Primates: key advances over the last two decades[J]. Journal of Neuroendocrinology, 2008, 20(6): 719-726.
[26]	VANSELOW J, FÜRBASS R. The bovine genome contains three differentially methylated paralogous copies of the P450c17 encoding gene (CYP17A1)[J]. General and Comparative End-ocrinology, 2011, 170(3): 475-479.
[27]	王霞. 藏绵羊睾酮合成相关基因在睾丸和附睾中的表达特征及生物学功能[D]. 兰州: 甘肃农业大学, 2021.
	WANG X. Expression characteristics and biological functions of testosterone synthesis-related genes in testis and epididymis of Tibetan sheep[D]. Lanzhou: Gansu Agricultural University, 2021.
[28]	ISRAEL J M, CABELGUEN J M, LE MASSON G, et al. Neonatal testosterone suppresses a neuroendocrine pulse gene-rator required for reproduction[J]. Nature Communications, 2014, 5: 3285.
[29]	AUCHUS R J. The genetics, pathophysiology, and management of human deficiencies of P450c17[J]. Endocrinology and Metabolism Clinics of North America, 2001, 30(1): 101-119, vii.
[30]	SMITH G C, SINCLAIR A H, NORMAN R J. Ontogeny of steroidogenesis in the fetal sheep gonad[J]. Endocrinology, 2001, 142(8): 3568-3576.
[31]	张艳. CYP17A1、CYP19A1基因变异与黔北麻羊产羔性状调控机制研究[D]. 贵阳: 贵州大学, 2022.
	ZHANG Y. Study on the regulatory mechanism of CYP17A1 and CYP19A1 gene variations and lambing traits in Guizhou Nor-thern Ma sheep [D]. Guiyang: Guizhou University, 2022
[32]	史东杰, 谭政, 张强, 等. 细胞色素CYP17a1基因在锦鲤性腺不同发育时期的表达分析[J]. 河南农业科学, 2022, 51(2): 139-146.
	SHI D J, TAN Z, ZHANG Q, et al. Expression of cytochrome CYP17a1 in different developmental phases of gonad in koi carp(Cyprinus carpio)[J]. Journal of Henan Agricultural Sciences, 2022, 51(2): 139-146.
[33]	杨晶, 郭晓飞, 张金龙, 等. 幼年与成年湖羊超数排卵及卵巢CYP17A1、INHA和FSHR基因表达差异研究[J]. 黑龙江畜牧兽医, 2021(9): 54-57.
	YANG J, GUO X F, ZHANG J L, et al. Study on the superovulation and expression difference of CYP17A1 INHA and FSHR genes in ovary between juvenile and adult Hu sheep[J]. Heilongjiang Animal Science and Veterinary Medicine, 2021(9): 54-57.
[34]	戴遥, 薛丽萍, 章诗琪, 等. CYP17A1基因突变致先天性肾上腺皮质增生症一例报道并文献复习[J]. 中国全科医学, 2025, 28(6): 771-776.
	DAI Y, XUE L P, ZHANG S Q, et al. Congential adrenal hyperplasia associated with CYP17A1 gene mutation: a case report and literature review[J]. Chinese General Practice, 2025, 28(6): 771-776.

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

物种名 Species name	NCBI登录号 NCBI login number	相似性 Similarity/%
山羊Capra hircus	NM_001314145.1	99.15
牛Bos taurus	NM_174304.3	96.93
白尾鹿Odocolieus virginianus	XM_020897894.2	96.27
猪Sus scrofa	NM_214428.1	85.62
驴Equus asinus	XM_014852085.3	82.94
马Equus caballus	NM_001082523.1	82.75
猫Felis catus	NM_001009371.2	82.75
犬Canis lupus familiaris	XM_038440431.1	81.76
智人Homo sapiens	NM_000102.4	80.26
兔Oryctolagus cuniculus	XM_002718585.5	77.91
鼠Mus musculus	NM_007809.3	72.09

氨基酸Amino acid	数量Quantity	比例Proportion/%
丙氨酸 Ala（A）	32	6.3
精氨酸 Arg（R）	22	4.3
天冬酰胺 Asn（N）	25	4.9
天冬氨酸 Asp（D）	24	4.7
半胱氨酸 Cys（C）	5	1.0
谷氨酰胺 Gln（Q）	23	4.5
谷氨酸 Glu（E）	30	5.9
甘氨酸 Gly（G）	27	5.3
组氨酸 His（H）	15	2.9
异亮氨酸 Ile（I）	36	7.1
亮氨酸 Leu（L）	70	13.8
赖氨酸 Lys（K）	36	7.1
蛋氨酸 Met（M）	10	2.0
苯丙氨酸 Phe（F）	27	5.3
脯氨酸 Pro（P）	28	5.5
丝氨酸 Ser（S）	35	6.9
苏氨酸 Thr（T）	23	4.5
色氨酸 Trp（W）	7	1.4
酪氨酸 Tyr（Y）	7	1.4
缬氨酸 Val（V）	27	5.3