Cloning,Bioinformatics and Tissue Expression Analysis of PFN3 Gene in Sangsang Yak from Tibet

doi:10.7668/hbnxb.20195594

Abstract

Abstract:

The aim of this study was to investigate the structure and physicochemical index of PFN3 gene(Profilin 3)in Sangsang yak,and to detect the expression of RPL8 gene in various tissues by qPCR,so as to lay a theoretical foundation for exploring the biological function of PFN3 gene in Sangsang yak.Using cardiac tissue cDNA as template,we cloned the CDS region of PFN3 gene in yak and detected its expression in different tissues.Bioinformatics analysis is performed using a variety of software and online tools.The results showed that the PFN3 coding region of Sangsang yak was 414 bp in length,with 3 open reading frames encoding 137 amino acids.The analysis of PFN3 protein in yak showed that the aliphatic coefficient was 101.75,the theoretical isoelectric point was 9.04,and the instability index was 37.22,indicating that PFN3 protein was a stable hydrophobic protein.The protein had 4 potential phosphorylation sites and no signal peptide and transmembrane region.Glycine(Gly)accounted for 14.6% of the total amino acids.Subcellular localization showed that PFN3 protein accounted for 43.5%,30.4% and 26.1% in cytoplasm,nucleus and mitochondria,respectively.Homology comparison showed that Sangsang yak had the closest homology with wild yak,and was the most closely related to African warthog and Eurasian boar.KEGG pathway enrichment analysis of interacting proteins showed that these proteins were mainly enriched in actin cytoskeleton regulation,Rap1 signaling pathway,focused adhesion,salmonella infection and amyotrophic lateral sclerosis.qPCR results showed that PFN3 gene expression in Sangsang yak was the highest in rectal tissue,significantly higher than that in other tissues,and the lowest in heart.

Key words: Sang mulberry yak, PFN3 gene, Clone, Bioinformatics and the analysis, Tissue expression

CLC Number:

S823.85

HE Wenbin, YANG Yan, Lobsong Dunzhu, LA Yongfu, REN Wenwen, MA Xiaoming, ZHANG Qiang, Zhuoma Tsering, Nima Gyatso, Phuntsok Zhandui, LIANG Chunnian. Cloning,Bioinformatics and Tissue Expression Analysis of PFN3 Gene in Sangsang Yak from Tibet[J]. Acta Agriculturae Boreali-Sinica, 2026, 41(2): 214-221. doi: 10.7668/hbnxb.20195594.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.20195594

http://www.hbnxb.net/EN/Y2026/V41/I2/214

Figures/Tables 9

References 26

[1]	李红梅, 包永清, 张红霞, 马登录, 旺青拉东. 美仁牦牛遗传资源特性分析与保护利用[J]. 畜牧兽医杂志, 2024, 43(5):118-121.doi:10.7606/j.issn.1004-6704.2024.05.026.
	Li H M, Bao Y Q, Zhang H X, Ma D L, Wang Q L D. Analysis conservation and utilization of genetic resources of Meiren yak[J]. Journal of Animal Science and Veterinary Medicine, 2024, 43(5):118-121.
[2]	李岩, 杨笑妍, 姜爱博, 祁霞, 牛小莹, 袁勇. 甘肃省牦牛产业发展现状及对策[J]. 甘肃畜牧兽医, 2024, 54(3):100-103,112.doi:10.15979/j.cnki.cn62-1064/s.2024.03.002.
	Li Y, Yang X Y, Jiang A B, Qi X, Niu X Y, Yuan Y. Development status and countermeasures of yak industry in Gansu Province[J]. Gansu Animal Husbandry and Veterinary Medicine, 2024, 54(3):100-103,112.
[3]	钟金城, 赵素君, 陈智华, 马志杰. 牦牛品种的遗传多样性及其分类研究[J]. 中国农业科学, 2006, 39(2):389-397.doi:10.3864/j.issn.0578-1752.at-2005-6068.
	Zhong J C, Zhao S J, Chen Z H, Ma Z J. Study on genetic diversity and classification of the yak[J]. Scientia Agricultura Sinica, 2006, 39(2):389-397.
[4]	何文斌, 扎西塔, 秦玉峰, 马金保, 赵敏慧, 孔祥颖, 胡林云, 梁春年. 放牧祁连牦牛屠宰性能及不同肌肉组织营养特性研究[J]. 中国草食动物科学, 2025, 45(5):98-103.doi:10.3969/j.issn.2095-3887.2024.00.116.
	He W B, Zha X T, Qin Y F, Ma J B, Zhao M H, Kong X Y, Hu L Y, Liang C N. Study on slaughtering performance of Grazing Qilian Yak and nutritional characteristics of different muscle tissues[J]. China Herbivore Science, 2025, 45(5):98-103.
[5]	马小勇, 陈育忠, 党智, 张贵男, 杨银芳, 王润丽, 牟艳, 庞生久, 何克磊, 王琴韦, 南青卓玛, 梁春年. 牦牛育肥影响因素的研究进展[J]. 中国草食动物科学, 2025, 45(2):95-99.doi:10.3969/j.issn.2095-3887.2024.00.123.
	Ma X Y, Chen Y Z, Dang Z, Zhang G N, Yang Y F, Wang R L, Mou Y, Pang S J, He K L, Wang Q W, Nan Q Z M, Liang C N. Research progress on influencing factors of Yak fattening[J]. China Herbivore Science, 2025, 45(2):95-99.
[6]	Gul S I, Ayoub A, Ali S A, Hanook S, Baig D N. Profilin 3 genetic architecture in glioma formalin fixed paraffin embedded(FFPE)archive[J]. Gene, 2021, 787:145614.doi:10.1016/j.gene.2021.145614. URL
[7]	李新荻, 王宇, 卢鸿, 罗布白珍, 何辰庆, 段梦琪, 叶幼荣, 张健, 商鹏. 猪PFN1基因在骨骼肌组织中的表达模式及多态性研究[J]. 家畜生态学报, 2023, 44(11):10-15.doi:10.3969/j.issn.1673-1182.2020.11.002.
	Li X D, Wang Y, Lu H, Luo B B Z, He C Q, Duan M Q, Ye Y R, Zhang J, Shang P. Expression pattern and polymorphism of pig PFN 1 gene in skeletal muscle tissue[J]. Journal of Domestic Animal Ecology, 2023, 44(11):10-15.
[8]	Behnen M, Murk K, Kursula P, Cappallo-Obermann H, Rothkegel M, Kierszenbaum A L, Kirchhoff C. Testis-expressed profilins 3 and 4 show distinct functional characteristics and localize in the acroplaxome-manchette complex in spermatids[J]. BMC Cell Biology, 2009, 10(1):34.doi:10.1186/1471-2121-10-34.
[9]	Tariq N, Basharat Z, Butt S, Baig D N. Distribution analysis of profilin isoforms at transcript resolution with mRNA-seq and secondary structure in various organs of Rattus norvegicus[J]. Gene, 2016, 589(1):49-55.doi:10.1016/j.gene.2016.05.023. URL
[10]	姚宝辉, 安康, 侯齐琪, 张德罡, 苏军虎. 高原鼢鼠精子发生相关基因的季节性表达特征[J]. 草原与草坪, 2024, 44(5):61-71.doi:10.13817/j.cnki.cyycp.2024.05.007.
	Yao B H, An K, Hou Q Q, Zhang D G, Su J H. Seasonal characteristics of spermatogenesis related gene expression in male plateau zokor[J]. Grassland and Turf, 2024, 44(5):61-71.
[11]	Hernández-Avilés C, Ramírez-Agámez L, Varner D D, Raudsepp T, Love C C. Spermatozoal acrosome dysfunction and its role in stallion subfertility[J]. Journal of Equine Veterinary Science, 2024, 143:105213.doi:10.1016/j.jevs.2024.105213. URL
[12]	Yaman O, Topcu-Tarladacalisir Y. L-carnitine counteracts prepubertal exposure to cisplatin induced impaired sperm in adult rats by preventing germ cell apoptosis[J]. Biotechnic & Histochemistry, 2018, 93(3):157-167.doi:10.1080/10520295.2017.1401661.
[13]	Fateen A, Ali S A, Baig D N, Basharat Z. Profilin(pfn)isoforms transcriptional and bioinformatic exploration and Mus musculus brain tissues development[J]. Gene Reports, 2018, 10:177-183.doi:10.1016/j.genrep.2017.12.005. URL
[14]	罗光荣, 肖敏, 何小强, 杨超, 李世林, 杨壮, 冯瑶. 牦牛产业发展新体系的思路与对策[J]. 当代畜牧, 2022(12):133-136.
	Luo G R, Xiao M, He X Q, Yang C, Li S L, Yang Z, Feng Y. Thoughts and countermeasures of the new system of yak industry development[J]. Contemporary Animal Husbandry, 2022(12):133-136.
[15]	徐宇辉, 吴志鹏, 盛欣, 李瑞哲, 马志杰. 牦牛全基因组关联研究进展[J]. 中国畜牧杂志, 2024, 60(12):41-47.doi:10.19556/j.0258-7033.20231221-10.
	Xu Y H, Wu Z P, Shen X, Li R Z, Ma Z J. Research progress on genome-wide association study in yak[J]. Chinese Journal of Animal Science, 2024, 60(12):41-47.
[16]	Suo L D, Gao M W, Ma T H, Gao Z M. Effect of RPL27 knockdown on the proliferation and apoptosis of human liver cancer cells[J]. Biochemical and Biophysical Research Communications, 2023, 682:156-162.doi:10.1016/j.bbrc.2023.10.012. URL
[17]	Umer N, Arévalo L, Phadke S, Lohanadan K, Kirfel G, Sons D, Sofia D, Witke W, Schorle H. Loss of Profilin3 impairs spermiogenesis by affecting acrosome biogenesis,autophagy,manchette development and mitochondrial organization[J]. Frontiers in Cell and Developmental Biology, 2021, 9:749559.doi:10.3389/fcell.2021.749559. URL
[18]	Sillero A, Ribeiro J M. Isoelectric points of proteins:theoretical determination[J]. Analytical Biochemistry, 1989, 179(2):319-325.doi:10.1016/0003-2697(89)90136-X. pmid: 2774179
[19]	田连琛, 原雅艺, 党旭红. 线粒体:电离辐射损伤的靶点[J]. 生物化学与生物物理进展, 2025, 52(4):836-844.doi:10.16476/j.pibb.2024.0394.
	Tian L C, Yuan Y Y, Dang X H. Mitochondria:the target of ionizing radiation damage[J]. Progress in Biochemistry and Biophysics, 2025, 52(4):836-844.
[20]	张慧, 黄吴键, 陈国勇, 梅晓妍, 谢鹏, 郑海英, 兰风华, 张朵. 精子鞭毛多发形态异常患者KLHL10基因突变分析[J]. 中国优生与遗传杂志, 2024, 32(7):1336-1342.doi:10.13404/j.cnki.cjbhh.2024.07.017.
	Zhang H, Huang W J, Chen G Y, Mei X Y, Xie P, Zheng H Y, Lan F H, Zhang D. Detection of KLHL10 gene mutations in patients with multiple morphological abnormalities of the sperm flagella[J]. Chinese Journal of Birth Health & Heredity, 2024, 32(7):1336-1342.
[21]	马小勇, 杨国武, 杨树猛, 丁考仁青, 唐月琴, 赵雪, 张艳丽, 梁春年. 美仁牦牛Akirin1基因克隆及在不同组织中的表达[J]. 甘肃农业大学学报, 2023, 58(6):20-30.doi:10.13432/j.cnki.jgsau.2023.06.003.
	Ma X Y, Yang G W, Yang S M, Ding K R Q, Tang Y Q, Zhao X, Zhang Y L, Liang C N. Cloning and expression of Akirin1 gene in different tissues of Meiren yak[J]. Journal of Gansu Agricultural University, 2023, 58(6):20-30.
[22]	张娟香, 马小勇, 路建卫, 仁青吉, 罗胜伟, 肖光峰, 王有鹤, 马晓明, 喇永福, 郭宪, 褚敏, 阎萍, 梁春年. 牦牛FTL基因克隆、生物信息学及表达分析[J]. 华北农学报, 2023, 38(6):192-200.doi:10.7668/hbnxb.20194239.
	Zhang J X, Ma X Y, Lu J W, Ren Q J, Luo S W, Xiao G F, Wang Y H, Ma X M, La Y F, Guo X, Chu M, Yan P, Liang C N. Cloning,bioinformatics analysis and expression analysis of FTL gene in yak[J]. Acta Agriculturae Boreali-Sinica, 2023, 38(6):192-200.
[23]	张城, 杨倩, 姚俊, 曹新. OSBPL2介导RhoA/ROCK2信号通路调控听觉细胞肌动蛋白细胞骨架[J]. 南京医科大学学报(自然科学版), 2023, 43(5):648-654.doi:10.7655/nydxbns20230509.
	Zhang C, Yang Q, Yao J, Cao X. OSBPL2 mediates RhoA/ROCK2 signaling pathway,regulating actin cytoskeleton in auditory cells[J]. Journal of Nanjing Medical University(Natural Sciences), 2023, 43(5):648-654.
[24]	靳英, 付小霞, 段瑞敏, 郝力瑶, 李峰. Rap1 GTP酶激活蛋白对结肠癌细胞增殖、侵袭和迁移作用的影响[J]. 临床与实验病理学杂志, 2024, 40(2):144-149,157.doi:10.13315/j.cnki.cjcep.2024.02.007.
	Jin Y, Fu X X, Duan R M, Hao L Y, Li F. Effects of Rap1 GTP-activating protein on proliferation,invasion and migration of colon cancer cells[J]. Chinese Journal of Clinical and Experimental Pathology, 2024, 40(2):144-149,157.
[25]	赖冬娣, 董涵, 魏亚明, 苑召虎. Sema7a促进内皮细胞与巨核细胞粘附的机制研究[J]. 临床输血与检验, 2024, 26(3):299-308.doi:10.3969/j.issn.1671-2587.2024.03.002.
	Lai D D, Dong H, Wei Y M, Yuan Z H. Exploration of the mechanism of Sema7a promoting adhesion of umbilical vein endothelial cells to megakaryocytes[J]. Journal of Clinical Transfusion and Laboratory Medicine, 2024, 26(3):299-308. doi: 10.3969/j.issn.1671-2587.2024.03.002
[26]	张永云, 赵跃, 王锐, 秦彩艳, 王配, 王淑燕, 李福泉, 刘丽仙, 霍海龙, 李卫真, 卢健, 霍金龙. 猪新基因PFN3电子克隆、RT-PCR验证和组织表达分析[J]. 云南农业大学学报:自然科学版, 2014, 29(2):179-186.doi:10.3969/j.issn.1004-390X(n).2014.02.006.
	Zhang Y Y, Zhao Y, Wang R, Qin C Y, Wang P, Wang S Y, Li F Q, Liu L X, Huo H L, Li W Z, Lu J, Huo J L. In Silico cloning,RT-PCR verification and tissues expression of novel swine gene PFN3[J]. Journal of Yunnan Agricultural University:Natural Science Edition, 2014, 29(2):179-186.

基因 Gene	用途 Purpose	序列(5'—3') Sequence (5'—3')	退火温度/℃ Annealing temperature	目的产物长度/bp Length of the target product
PFN3	PCR	F1 ACTTCATAACCACGCCCCTG	57	567
		R1 CACGCCCAGGTCAGACTTTA
PFN3	qPCR	F2 TGGGCGACTGGAAAGGTTAC	58	82
		R2 TTGTCCGAGTGGCCCACAAT
GAPDH	qPCR	F2 TTGTGATGGGCGTGAACC	58	169
		R2 GTCTTCTGGGTGGCAGTGAT

基因 Gene	用途 Purpose	序列(5'—3') Sequence (5'—3')	退火温度/℃ Annealing temperature	目的产物长度/bp Length of the target product
PFN3	PCR	F1 ACTTCATAACCACGCCCCTG	57	567
		R1 CACGCCCAGGTCAGACTTTA
PFN3	qPCR	F2 TGGGCGACTGGAAAGGTTAC	58	82
		R2 TTGTCCGAGTGGCCCACAAT
GAPDH	qPCR	F2 TTGTGATGGGCGTGAACC	58	169
		R2 GTCTTCTGGGTGGCAGTGAT

软件 Software	用途 Application
NCBI BLAST(https://blast.ncbi.nlm.nih.gov/Blast.cgi)	序列对比验证
MEGA 10.1	构建系统进化树
Open reading frame finder(OFR Finder, http://www.ncbi.nlm.nih.gov/gorf/orfig.cgi)	查看开放阅读框
ProtParam(http://www.expasy.ch/tools/Protparam.html)	蛋白质理化性质分析
ProtScale(https://web.expasy.org/protscale/)	蛋白亲疏水性分析
SignalP 4.1(http://www.cbs.dtu.dk/services/SignalP/)	信号肽预测
TMHMM(http://www.cbs.dtu.dk/services/TMHMM/)	跨膜结构域分析
Psort Ⅱ(https://wolfpsort.hgc.jp/)	蛋白亚细胞定位
NetPhos 3.1 Server(http://www.cbs.dtu.dk/services/NetPhos/)	磷酸化位点预测
Sopma(https://npsa-prabi.ibcp.fr/cgi- bin/npsa_automat.pl?page=npsa_sopma.html)	蛋白质二级结构预测
SWISS-MODEL(http://www.expasy.org/swissmod/SWISS-MODEL.html)	蛋白质三级结构预测
STRING(https://string-db.org/)	蛋白互作网络预测

软件 Software	用途 Application
NCBI BLAST(https://blast.ncbi.nlm.nih.gov/Blast.cgi)	序列对比验证
MEGA 10.1	构建系统进化树
Open reading frame finder(OFR Finder, http://www.ncbi.nlm.nih.gov/gorf/orfig.cgi)	查看开放阅读框
ProtParam(http://www.expasy.ch/tools/Protparam.html)	蛋白质理化性质分析
ProtScale(https://web.expasy.org/protscale/)	蛋白亲疏水性分析
SignalP 4.1(http://www.cbs.dtu.dk/services/SignalP/)	信号肽预测
TMHMM(http://www.cbs.dtu.dk/services/TMHMM/)	跨膜结构域分析
Psort Ⅱ(https://wolfpsort.hgc.jp/)	蛋白亚细胞定位
NetPhos 3.1 Server(http://www.cbs.dtu.dk/services/NetPhos/)	磷酸化位点预测
Sopma(https://npsa-prabi.ibcp.fr/cgi- bin/npsa_automat.pl?page=npsa_sopma.html)	蛋白质二级结构预测
SWISS-MODEL(http://www.expasy.org/swissmod/SWISS-MODEL.html)	蛋白质三级结构预测
STRING(https://string-db.org/)	蛋白互作网络预测

氨基酸 Amino acid	数量/个 Quantity	含量/% Content	氨基酸 Amino acid	数量/个 Quantity	含量/% Content
丙氨酸Ala (A)	11	8.0	亮氨酸Leu (L)	16	11.7
精氨酸Arg (R)	14	10.2	赖氨酸Lys (K)	3	2.2
天冬酰胺Asn (N)	2	1.5	蛋氨酸Met (M)	3	2.2
天冬氨酸Asp (D)	10	7.3	苯丙氨酸Phe (F)	1	0.7
半胱氨酸Cys (C)	5	2.9	脯氨酸Pro (P)	4	2.9
谷氨酰胺Gln (Q)	4	2.9	丝氨酸Ser (S)	6	4.4
谷氨酸Glu (E)	3	2.2	苏氨酸Thr (T)	6	4.4
甘氨酸Gly (G)	20	14.6	色氨酸Trp (W)	2	1.5
组氨酸His (H)	6	4.4	酪氨酸Tyr (Y)	1	0.7
异亮氨酸Ile (I)	8	5.8	缬氨酸Val (V)	12	8.8

Please choose a citation manager

Content to export