基于PRLR-JAK-STAT5信号传导通路测定鹅催乳素的新方法

doi:10.11843/j.issn.0366-6964.2016.07.011

Abstract

Abstract:

In order to develop a high sensitive method of detecting goose prolactin，luciferase detecting system based on PRLR-JAK-STAT5 signal transduction pathway was designed in the present study.Firstly the CDS region of goose PRLR gene was cloned，the signal transducer and activator of transcription 5（STAT5) was artificially synthesized and inserted into the eukaryotic expression vector pCMV6-Entry and luciferase reporter pGL3-Enhancer plasmid as signal receiver vector and signal responder vector respectively.Secondly，the two recombinant vectors along with puromycin screening vector pEZX-MR03 and internal control pRL-TK vector were transfected into HEK293T cells.After puromycin screening，stable transgenic cell lines were isolated and stimulated by a serial of different concentrations of chicken PRL (0，30，60，90 ng•mL-1).Finally the mRNA relative expression level and enzyme activity of luciferase enzyme were detected by qRT-PCR and dual-luciferase detection system.Results showed that 10 stable transgene cell lines carried the 4 vectors were isolated.After treated with different concentrations of chicken PRL，luciferase gene mRNA expression level was up regulated and luciferase enzyme activity also enhanced in dose dependent manner in one of these 10 cell lines.These results demonstrated that it was feasible of detecting PRL bioactivity using the constructed PRLR-JAK-STAT5 signal transduction system and could be further used in measuring poultry PRL concentrations.

CLC Number:

S835.2

SONG Jin-wei，LI Hui，CHEN Zhe，SHI Zhen-dan，WANG Zhen-yong. A Novel Method of Detecting Goose Prolactin Based on PRLR-JAK-STAT5 Signal Transduction Pathway[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(7): 1389-1395.

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References

［1］王锋.动物繁殖学［M］.北京：中国农业大学出版社，2012：53-54.
WANG F.Animal reproduction science［M］.Beijing：China Agricultural University Press，2012：53-54.(in Chinese)
［2］李静，滕卫平.催乳素的免疫调节作用［J］.国外医学内分泌学分册，2000，20(2)：99-101.
LI J，TENG W P.Immune regulation of prolactin［J］.Foreign Medical Sciences Endocrine Section，2000，20(2)：99-101.(in Chinese)
［3］施振旦，陈峰，毕英佐.禽类就巢发生和调控研究进展［J］.黑龙江动物繁殖，2000，8(3)：37-41.
SHI Z D，CHEN F，BI Y Z.Research on regulation of poultry seasonal reproduction［J］.Heilongjiang Journal of Animal Reproduction，2000，8(3)：37-41.(in Chinese)
［4］刘颖，施振旦，黄运茂，等.鸡催乳素和抑制素-α亚基重组蛋白的构建及表达［J］.中国兽医学报，2002，22(4)：365-367.
LIU Y，SHI Z D，HUANG Y M，et al.Construction and expression of recombinant proteins based on chicken prolactin and inhibin α subunit fragment［J］.Chinese Journal of Veterinary Science，2002，22(4)：365-367.(in Chinese)
［5］杨海明.扬州鹅繁殖的光照调控及下丘脑基因差异表达研究［D］.扬州：扬州大学，2014.
YANG H M.Study on light regulation and differential expression of genes in hypothalamus during Yangzhou geese reproductive cycle［D］.Yangzhou：Yangzhou University，2014.(in Chinese)
［6］黄运茂，施振旦.广东地方鹅产蛋—就巢周期调控模式的探讨［J］.黑龙江畜牧兽医，2007(7)：57-58.
HUANG Y M，SHI Z D.Study on the regulation pattern of Guangdong local goose seasonal reproduction［J］.Heilongjiang Animal Science and Veterinary Medicine，2007(7)：57-58.(in Chinese)
［7］施振旦，朱基美，黄运茂，等.调节马岗鹅繁殖特性的初步结果和结论［J］.中国草食动物，2001，3(1)：12-15.
SHI Z D，ZHU J M，HUANG Y M，et al.Preliminary results and conclusions of the regulation of the reproductive characteristics of Magang goose［J］.China Herbivores，2001，3(1)：12-15.(in Chinese)
［8］施振旦，黄祖汉.家鸡催乳素放射免疫测定法的建立［J］.核农学报，2000，14(1)：36-39.
SHI Z D，HUANG Z H.Using micro-quantity of chloraminet in chicken prolactin labelling and radioimmunoassay［J］. Acta Agriculturae Nucleatae Sinica，2000，14(1)：36-39.(in Chinese)
［9］DAWSON A，SHARP P J.The role of prolactin in the development of reproductive photorefractoriness and postnuptial molt in the European starling (Sturnus vulgaris)［J］.Endocrinology，1998，139(2)：485-490.
［10］SHI Z D，HUANG Y M，LIU Z，et al.Seasonal and photoperiodic regulation of secretion of hormones associated with reproduction in Magang goose ganders［J］.Domest Anim Endocrinol， 2007，32(3)：190-200.
［11］HUANG Y M，SHI Z D，LIU Z，et al.Endocrine regulations of reproductive seasonality，follicular development and incubation in Magang geese［J］.Anim Reprod Sci，2008，104(2-4)：344-358.
［12］郭日红.鹅催乳素ELISA分析方法的建立［D］.广州：华南农业大学，2013.
GUO R H.Devlopment of enzyme-linked immunoadsorbent assay (ELISA) for measuring goose prolactin［D］.Guangzhou：South China Agriculture University，2013.(in Chinese)
［13］YAMASHITA S，TAKAYANAGI A，SHINIZU N.Temporal and cell-type specific expression of c-fos and c-jun protooncogenes in the mouse uterus after estrogen stimulation［J］.Endocrinology，1996，137(12)：5468-5475.
［14］GOFFIN V，FERRAG F，KELLY P A.Chapter 1 Molecular aspects of prolactin and growth hormone receptors［J］.Adv Mol Cell Endocrinol，1998，2：1-33.
［15］周永明，田胜利，周韶虹，等.骨髓增生异常综合征患者JAK2和STAT5基因表达的变化［J］.临床血液学杂志，2007，20(2)：70-72.
ZHOU Y M，TIAN S L，ZHOU S H，et al.The comparative study on JAK2/STAT5 signal transduction pathway in myelodysplastic syndrome［J］.Journal of Clinical Hematology，2007，20(2)：70-72.(in Chinese)
［16］孙胜利.脑缺血和细胞缺氧/复氧过程中JAK-STAT信号转导相关基因的表达［D］.上海：第二军医大学，2006.
SUN S L.JAK/STAT-related gene expression during transient focal cerebral ischemia/reperfusion and during cell hypoxia/reoxygenation［D］.Shanghai：Second Military Medical University，2006.(in Chinese)
［17］SCHINDLER C，DARNELL J E.Transcriptional responses to polypeptide ligands：the JAK-STAT pathway［J］.Annu Rev Biochem，1995，64：621-651.
［18］ADACHI H，TAKEMOTO Y，BUNGO T，et al.Chicken leptin receptor is functional in activating JAK-STAT pathway in vitro［J］.J Endocrinol，2008，197(2)：335-342.
［19］ROSENBLUM C I，TOTA M，CULLY D，et al.Functional STAT 1 and 3 signaling by the leptin receptor (OB-R)；reduced expression of the rat fatty leptin receptor in transfected cells［J］.Endocrinology，1996，137(11)：5178-5181.
［20］ROSENBLUM C I，VONGS A，TOTA M R，et al.A rapid，quantitative functional assay for measuring leptin［J］.Mol Cell Endocrinol，1998，143(1-2)：117-123.
［21］HEN G，YOSEFI S，RONIN A，et al.Monitoring leptin activity using the chicken leptin receptor［J］.J Endocrinol，2008，197(2)：325-333.
［22］OHKUBO T，HIROTA K，MURASE D，et al.Avian blood induced intranuclear translocation of STAT3 via the chicken leptin receptor［J］.Comp Biochem Physiol B Biochem Mol Biol， 2014，174：9-14.

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A Novel Method of Detecting Goose Prolactin Based on PRLR-JAK-STAT5 Signal Transduction Pathway

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