CN115804361B 一种培育高雄性尼罗罗非鱼的方法 （山东农业大学）

本发明公开了一种培育高雄性尼罗罗非鱼2所述Kdm6bb_tv1基因的核苷酸序列如SEQIDNO.2.含有Kdm6bb_tv1基因的重组表达载体在提高高尼罗罗非鱼的雄性率是指提高Kdm6bb_tv1过表达鱼交配后代组表达载体是将Kdm6bb_tv1基因插入到pT2AL200R150G3鱼的重要养殖种类，吉富罗非鱼(GeneticImprovementofFarmedTilapia)是一个人工[0004]尼罗罗非鱼是具有XX/XY性别决定系统的雌雄异体鱼类，尼罗罗非鱼的性别分化受遗传和温度双重调控，属于遗传+温度性别决定(GeneticSexDeterminationplus利用鉴定出的新基因靶向性的培育高雄性率的转基因罗非鱼，将具有十分重要的应用价4[0017](3)将F1代个体性成熟后同型交配获得F2代，从F2代中筛选出阳性个体，获得[0023](1)本发明发现Kdm6bb_tv1是尼罗罗非鱼的雄性性别决定基因；通过过表达[0024](2)Kdm6bb_tv1基因过表达与高温辅助培育相结合具有协同促进作用，能够大幅[0028]说明高温36℃处理或者过表达Kdm6bb_tv1能诱导XX遗传型雌鱼性腺发育为精巢，5[0033]本发明发现尼罗罗非鱼内源的Kdm6bb_tv1是尼罗罗非鱼的雄性性别决定基因，[0034]atgtatcactcagtagagctctactctggccgtaacacatgggactcctactcagttggaggacctaacaaaggacaatgggctcctgtaaatgctcgcccctggagccacacacagaggctgcaagaagggcgcaaacagccatatcattctctgtcaagtcgtatttatagcaggggtgacagaacaatcaaccatgtcccggacaaaggtatctcaaaggggcatagccagctgctgcgaatttgggatggtaaagagctgcagtttgaggtccagaactggcatcacaactccatccgttcgttccacatccgagatggcaccaacaatggttatctgccaggaccagaggaacgctacgcaaactggcaaaacaataacgggggccctcgcctgcatcgcaacaacagggagcttcagggaaccgcgccagagagatgggcccactcagacccccgcaggagtttcccagacaggatggtgaacaacagaacaggaccatggaaacggccagccctccaccagcgacgggagcaagcgcatcaccacagtccacccccggagcacccattaaccccccgggatgagtgtccagctaagaggagaagagactcgggacctgatcaggcttgtcatactggatccaggcacttacctttacttgcccatgccccatcatcacctcaccaacagcgctccaaccaggatgattggaagcctccaagtgacagggcgggtccatgccaccactctgaacacaggacctcaacaacacagcaacaggaaacctctaaactgcgagctgggagacacggcttcagtgacccagtgccgtccaaccagagctgcggcaggaggccaccacattatggaagcagagggaaggtcgaacggaaaatctcatcttcaccagcagaccacacccgtgtgccttacagccaccaacaccatcactataaccatcaacggcacacaggccagcctagagcacctcagcacaacgccccactacaccctctggaggataaggactcttggaaccatcatcacaaacaaagcacagagaatcagcgcattcatcaacgaacaaattcaagggacccctccctctccaaatctggtgtagactctcctcagtactcatctgtcctctgtggtccgaacaatggaggttctacttccagcagtacaggtgctccttccagtccttcttcagacaaaatgctcaatggcagaaaagattcaccagtcattcagcagcgttgtcccagcctcagtgggcagctaaagctccaagaaagtcccagtgtcattccgagacccagtcgggcatcaagtgcatctccgacaccccgtgctggttcaaaatctaggctttcagacctcaagtacaggaagacctcacagctcctctgctcacggcagtctccccacgatagatcaagagtggacaagcctgagagggagttagaagataaaaagtctgaatccaaacaaaagtataagctgctgaaaaaggagaagaagggtgacgttctgaagatgggcagaaagggtgaggacagaaaaaggcggaagaaaaaagaggagaaaaggttgaccgagaagaaaaagaaacgcgataaagcatctaagaaggaacgaaaattagccctcaaaatgaaagttacagaacaggaaatgttttcttccgtctctgcttccacctttctaagtgaggccaagataagtaaaatggagactttgaacggagagaatcaggcccagtcaccacccaaacacaagcacagggacaggagtgaaaaaggagagaagactacgcataggtcatccacaaatacccttcatcccaggcccactttacagcagtcatcctcaaagtctgaaaatcacaaaatgctcaaaaagagcagcagtgccccagaaacccctcttcctaagaaaaccagccaccaggccagcaagcgttccaccatcgtctcaacccaactggaaggtgagccaaaagcaaagccagatgacacactcccttccctcttatttaaagccttagcgcctctcagtacagtgtgctcagctggcttagagcagcaaggcgggcagggaggagttctcaacgctccagaccttcagcccgtggctgtgatgggaaatttaagggaaacgggagacaaccttgccaacacacctcctgtgcttagctggcagggatctccagtatcagatcttggagatgatgaggaggagctagagaaaggtgtactaagcagacccgtcctccagctcagccccactcaatgtctttcaccccctcctgtagagagtgaaagcattgataatatgaatgcagaagattattcccacagcagcatgtctgaattctccgatctaccttgttcaattgaacaagtttcggaggaagaaaaggaagtgatgatggacagcagagggaagctgtctggctctccctttcatgagtttcataacgacgacacag6gttttgatgatgtctttgagagcctggccaccttgcttgccagccagagggtcgcgtgccgaggtggtccattcggcggtcctcctgctagtgatgcaaatggagtaaagtactcttgctctcttgaactgggcccagatatccactgccccgagaatcaggacttcttccctacatcatatcccacagaatcctccgaacccagtaatcaatcaccagttcacattaccacagacacacttgaagaattccacagtctggcagatgtgagtaagccagccaccagctcaatggtgcaggaaaaagaggaggaaaatgagagtgatgtacaaggtgcgatgaaacaagacggcaaggatgcagcaatcacgatggagaggaaggagactcttttggatgggtcattgagtgcagagctgactctgaccacaacacatacagtaatgaagatggtaaagaaagctccctctaccaaggaaaaaagtggaaagggcacaaagacagaacgtgcaggcaaagagaaaaagagaaaacagaaggtaaaggatgggaaaggagagggagcaatcaagattaagataaagagagaagaaaataaggtcatcagctgtaaaattaaaacaagtagagtacctgatgttaaagacagcgacacttcatcttcagtgccagttatttcatactcagccagaccggtcaaagacagcatgaagggccagactcctcaggaaaatcaaaccccaaatggcaaaaatatcaaaagacagaaaatggacactgggaattcaaatgcaacaacagatgagaagaaagggttagataatgagactgcctgtgtagtcggaattacagacatcaccacctcaaattcagcatcagctgtatcaacaagtccatccaaattagatcgtcctacaccagcaagtaaagtggacccactgaaactgaaagcgctgtccatgggcttgtctaaggagctgaagatcctcttggttaaagtggagagcggcggaaggcagacattcaacatatcagaggtgcaggagcaacgaatcccaatcccactttccaagatcagcatcacaaatacagccgctgatgtgatcggagcatgcaggggggcaagtgtgaaagggaaattcaaggagtcgtacttgcctcccgcattttcggttaaacccaacattgcagctgagacccccattcctcgcgataagctgaatcctcctacaccaagcatctatttggagagcaagagggatgccttttctccggttctgcttcagttctgcactgatcccaaaaacgctgttacggtcatcagaggcctcgctggctccctccgccttaatcttggtctgttctcaaccaaatctctggtggaggccaattcggaccatgcagtggaagtgaggactcaggttcagcagcctgctgatgagaactgggatccaagtggtgcgactcagacgtggccctgcgagagcagccgctcacacaccaccattgccaaatatgcccagtaccaggcctccacctttcaggagagcctggaggaggagaaggagagtgagaatgaggaagacgaggaacagactccaaaccaatcggcgtccacaaaagccgctctgacattatccaacagtaaaggcaatcatacctcaaaagccagttctgcgtccatcttgagcaaagctcagcctcctaatgccaacagctctctgagctcagagcaaaaacctgttggaaagatcattaaattcgggaccaacatagatctgtctgatcctaaaaggtggaagccccagctgcaggagctgctgaagctgccagctttcatgcgagtggaatccagcaacaacatgctgagtcacgtcggtcacaccatcctgggcatgaacactgtccagctttacatgaaggtgccaggcagccgtacgccaggtcatcaagagaacaacaatttctgctcagttaacatcaacatcgggcccggtgactgtgagtggttcgcagtccatgaacactactgggaacttattaacaatttatgtgaaaagcatggagtagactaccttacagggtcctggtggccagttctagaggatctctacagttccaacatccctgtgtaccgcttcatccagaggccaggcgacctggtgtggattaatgcaggaactgtgcactgggtccaagctgtgggctggtgcaacaacattgcctggaatgtgggaccgctcaactcataccaatatcaactcgccctggagcgctttgagtggaacgaggtgaagaaggttaagtcaatcgttcccatgatccacgtttcctggaatgtggctcgcaccattaaaatcacagatcaggataccttcaagatgatcaaacactgcctgttgcagtccatcaagcacatccagattctgagagaccagctggtggctgcagggaagaaaatctgttaccagagccgcgtgaaagacgagccagcctactactgcaacgagtgtgatgtggaggtgtttaacttgctgtttgtgacaagtgagaacagtagtaagaagacctacgtggtgcactgtgaggactgtgccagagctaagaacccgtcgctgacaggagtagtggtgctggaacagtatcggatagaggagctgatgaaaatctatgacagttttgtgctgactccaactccct7[0038](2)Kdm6bb_tv1过表达载体的构建：将上述的Kdm6bb_tv1基因片段插入一种转基体和体外转录合成的转座酶mRNA(pCS_TP质粒为模板，体外转录合成的)共注射到1细胞期[0040](5)Kdm6bb_tv1过表达F2个体的饲养及雄性率统计：Kdm6bb_tv1过表达F2代个体28℃水温继续饲养至性成熟。[0043]说明高温36℃处理或者过表达Kdm6bb_