华北农学报
華北農學報
화북농학보
ACTA AGRICULTURAE BOREALI-SINICA
2013年
5期
6-14
,共9页
李红霞%李建林%唐永凯%俞菊华%阮瑞霞%魏可鹏%夏正龙%董在杰
李紅霞%李建林%唐永凱%俞菊華%阮瑞霞%魏可鵬%夏正龍%董在傑
리홍하%리건림%당영개%유국화%원서하%위가붕%하정룡%동재걸
建鲤%生长激素促泌素受体%转录变体%表达
建鯉%生長激素促泌素受體%轉錄變體%錶達
건리%생장격소촉비소수체%전록변체%표체
Cyprinus carpio var.jian%Growth hormone secretagogue-receptor%Transcript variants%Expression
应用RT-PCR及PCR分离到鲤鱼2个jlGHSR2s基因，jlGHSR2a和2b，分别编码366，367个氨基酸，相似性高达98％，和金鱼、斑马鱼GHSR2的相似性高达95％。 jlGHSR2s基因的内含子位于起始密码子793 bp后，2a和2b内含子的长度和序列均存在明显差异。试验首次发现了jlGHSR2 a的转录变体（Variant）jlGHSR2a′，由选择性剪切靠近内含子的57 bp外显子1和保留靠近外显子2的28 bp内含子构成，编码350个氨基酸，缺失最后2个跨膜区，剪切序列两端碱基为CT-AC。荧光定量RT-PCR结果表明，jlGHSRs在脑和精巢表达量最高，1s在检测的组织均有表达，但2s在其他组织表达量很少。 jlGHSRs的转录变体1s′和2a′的表达量与1s和2s有一定的正相关性，但1s/1s′在各组织不一致，比值最高出现在肝脏，其次是肌肉、肠和脑，精巢和卵巢最低。生长对脑中jlGHS-R1s的表达量没有显著影响，但生长慢的个体肝和肠中jlGHSR1s的表达量比生长快的高，1s/1s′也有明显升高；肌肉中则相反，1s/1s′在雄鱼肌肉中减小明显。饥饿使雌鱼脑、雌鱼前肠和雌雄鱼后肠jlGHSR1s的表达量显著降低，对其余组织中的表达则不显著。生长快慢或饥饿对脑中jlGHSR2s的表达量均没有显著影响。结果表明，jlGHSR1s的表达量与生长有明显的相关性。
應用RT-PCR及PCR分離到鯉魚2箇jlGHSR2s基因，jlGHSR2a和2b，分彆編碼366，367箇氨基痠，相似性高達98％，和金魚、斑馬魚GHSR2的相似性高達95％。 jlGHSR2s基因的內含子位于起始密碼子793 bp後，2a和2b內含子的長度和序列均存在明顯差異。試驗首次髮現瞭jlGHSR2 a的轉錄變體（Variant）jlGHSR2a′，由選擇性剪切靠近內含子的57 bp外顯子1和保留靠近外顯子2的28 bp內含子構成，編碼350箇氨基痠，缺失最後2箇跨膜區，剪切序列兩耑堿基為CT-AC。熒光定量RT-PCR結果錶明，jlGHSRs在腦和精巢錶達量最高，1s在檢測的組織均有錶達，但2s在其他組織錶達量很少。 jlGHSRs的轉錄變體1s′和2a′的錶達量與1s和2s有一定的正相關性，但1s/1s′在各組織不一緻，比值最高齣現在肝髒，其次是肌肉、腸和腦，精巢和卵巢最低。生長對腦中jlGHS-R1s的錶達量沒有顯著影響，但生長慢的箇體肝和腸中jlGHSR1s的錶達量比生長快的高，1s/1s′也有明顯升高；肌肉中則相反，1s/1s′在雄魚肌肉中減小明顯。饑餓使雌魚腦、雌魚前腸和雌雄魚後腸jlGHSR1s的錶達量顯著降低，對其餘組織中的錶達則不顯著。生長快慢或饑餓對腦中jlGHSR2s的錶達量均沒有顯著影響。結果錶明，jlGHSR1s的錶達量與生長有明顯的相關性。
응용RT-PCR급PCR분리도리어2개jlGHSR2s기인，jlGHSR2a화2b，분별편마366，367개안기산，상사성고체98％，화금어、반마어GHSR2적상사성고체95％。 jlGHSR2s기인적내함자위우기시밀마자793 bp후，2a화2b내함자적장도화서렬균존재명현차이。시험수차발현료jlGHSR2 a적전록변체（Variant）jlGHSR2a′，유선택성전절고근내함자적57 bp외현자1화보류고근외현자2적28 bp내함자구성，편마350개안기산，결실최후2개과막구，전절서렬량단감기위CT-AC。형광정량RT-PCR결과표명，jlGHSRs재뇌화정소표체량최고，1s재검측적조직균유표체，단2s재기타조직표체량흔소。 jlGHSRs적전록변체1s′화2a′적표체량여1s화2s유일정적정상관성，단1s/1s′재각조직불일치，비치최고출현재간장，기차시기육、장화뇌，정소화란소최저。생장대뇌중jlGHS-R1s적표체량몰유현저영향，단생장만적개체간화장중jlGHSR1s적표체량비생장쾌적고，1s/1s′야유명현승고；기육중칙상반，1s/1s′재웅어기육중감소명현。기아사자어뇌、자어전장화자웅어후장jlGHSR1s적표체량현저강저，대기여조직중적표체칙불현저。생장쾌만혹기아대뇌중jlGHSR2s적표체량균몰유현저영향。결과표명，jlGHSR1s적표체량여생장유명현적상관성。
Growth hormone secretagogue receptors ( GHSRs ) , the endogenous receptor of growth hormone se-cretion ( ghrelin) ,belong to G-protein-coupled receptors family ,and play a regulatory role on animal growth ,energy homeostasis etc.In this experiment,RT-PCR and PCR methods were applied to isolate two jlGHSR2s,jlGHSR2a and 2b,eccoding 366 and 367 amino acids sharing 98% identity with each other,and 95% identity with Danio rerio GHSR2.Intron of jlGHSR2s is located at 793 bp after start codon,there are obvious differences in length and se-quences between 2a and 2b introns.We first report jlGHSR2a′,the variant of jlGHSR2a,which is selectively spliced last 57 bp of exon 1 and reserved last 28 bp of intron near exon 2.jlGHSR2a′encodes 350 amnio acids,missing the last two transmembrane region ,and two ends of splicing sequence are CT-AC.Fluorescent quantitative RT-PCR re-sults showed that the highest expression level of jlGHSRs is in brain and testis ,1s is expressed in all the tissues ex-amined in this experiment ,but2 s is rarely expressed in other tissues .The expression of variants 1 s′and 2 a′has cer-tain degree of positive correlation with 1s and 2s,but 1s/1s′is inconsistent among these tissues ,the highest ratio is Growth hormone secretagogue receptors ( GHSRs ) , the endogenous receptor of growth hormone se-cretion ( ghrelin) ,belong to G-protein-coupled receptors family ,and play a regulatory role on animal growth ,energy homeostasis etc.In this experiment,RT-PCR and PCR methods were applied to isolate two jlGHSR2s,jlGHSR2a and 2b,eccoding 366 and 367 amino acids sharing 98% identity with each other,and 95% identity with Danio rerio GHSR2.Intron of jlGHSR2s is located at 793 bp after start codon,there are obvious differences in length and se-quences between 2a and 2b introns.We first report jlGHSR2a′,the variant of jlGHSR2a,which is selectively spliced last 57 bp of exon 1 and reserved last 28 bp of intron near exon 2.jlGHSR2a′encodes 350 amnio acids,missing the last two transmembrane region ,and two ends of splicing sequence are CT-AC.Fluorescent quantitative RT-PCR re-sults showed that the highest expression level of jlGHSRs is in brain and testis ,1s is expressed in all the tissues ex-amined in this experiment ,but2 s is rarely expressed in other tissues .The expression of variants 1 s′and 2 a′has cer-tain degree of positive correlation with 1s and 2s,but 1s/1s′is inconsistent among these tissues ,the highest ratio is <br> in the liver,followed by muscle,intestine and brain,testis and ovary are the minimum ones .Growth does no signifi-cant effect on the jlGHS R1s expression in brain,but jlGHSR1s expression in liver and intestine of slow growth is higher than fast growth ones, 1s/1s′is also increased obviously.It is opposite in muscle,1s/1s′in male muscle is obviously decreased.The growth speed or starvation could do no significantly effects on jlGHSR2 s expression in the brain.Our results show that the jlGHSR1s expression level is clearly correlated with growth .
