Molecular Cloning and Expression Analysis of a AGAMOUS-like 66 Gene (GbAGL66) in Ginkgo biloba

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2 OPEN ACCESS Biotechnology ISSN X DOI: /biotech Research Article Molecular Cloning and Expression Analysis of a AGAMOUS-like 66 Gene (GbAGL66) in Ginkgo biloba Jinshuang Dou, Lanlan Wang, Jiaping Yan, Mingyue Fu, Xian Zhang and Feng Xu College of Horticulture and Gardening, Yangtze University, Jingzhou, Hubei, China

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5 1 CATCCCCTAAAAATCTGTCGGTACAGAAAACCATGGGCAGAGTTAAGCTCCCGATTAAGAAAATAGAGAATAGCA 1 M G R V K L P I K K I E N S 76 CTAACAGGCAGGTCACCTTCTCCAAACGGAGAAATGGATTGATCAAAAAGGCATATGAACTCTCAGTTCTGTGCG 25 T N R Q V T F S K R R N G L I K K A Y E L S V L C 151 ACATCGAAATTGCACTCATAATGTTCTCGCCCTCGGGAAGATTGAGCCACTTTTCAGGGAAAAATAGTAGGATAG 50 D I E I A L I M F S P S G R L S H F S G K N S R I 226 AAGATGTTATAGCTCGCTTCGTCAATCTGCCTGAGCACGAGAGGCCAAGGCTTGTCCAAAACCAAGAATATCTCC 75 E D V I A R F V N L P E H E R P R L V Q N Q E Y L 301 TCAGAGCGTTGAAAAAACTCAAGTACGAAAGCGATATCGCCAATCATCTTGCAAGCCCAAACATTGTCGACTCAA 100 L R A L K K L K Y E S D I A N H L A S P N I V D S 376 ATGTGGAGGAACTTCAAATGGATATTCGGAGACAGAGAATTCAGTTGGAGGAAGCCCAGCAGAAGTTAAGGAGTT 125 N V E E L Q M D I R R Q R I Q L E E A Q Q K L R S 451 TTAAAGAGGATCCACTTCTTATAACCTCAATACAAGATGCAGATCAATATGAAAGGACATTGGAGGAGGCTTTGC 150 F K E D P L L I T S I Q D A D Q Y E R T L E E A L 526 GTAGAGTTCGTCTCAGAAAACAACAGCTAGAACACAACCAAATGGCAGTTGCTTTCAACGATGCAAACTTGCAGT 175 R R V R L R K Q Q L E H N Q M A V A F N D A N L Q 601 TTTATATACAAACACAGAATGGATTGCCAAATGGAACAGACACAAGTCAAAATCACCTATACAACTCATGGATGC 200 F Y I Q T Q N G L P N G T D T S Q N H L Y N S W M 676 CACAGGGAGACCCCCATACTAGTGTCCAGAATTTTATGGAGCATGAAAATTCTAATGCCATGCTTGCGATGCGGG 225 P Q G D P H T S V Q N F M E H E N S N A M L A M R 751 AAGCACAATGCATGGCAAAGTGTTTACAAAATGGAACCGTATTTCCGGCACTTCAAGATGCCACTGGAATGCAGC 250 E A Q C M A K C L Q N G T V F P A L Q D A T G M Q 826 TTCCAAATGAGTCTGCAAGTACTCAACCGTATATTCCTACATCACACATGCAGTTTGATTACACTTTAACTGACA 275 L P N E S A S T Q P Y I P T S H M Q F D Y T L T D 901 ACAACAATAATGAACATGCAGAACAGGCCGATATAGCTGCAGCGTTTGACTATGGTTCTGATGCAATGGCATCCG 300 N N N N E H A E Q A D I A A A F D Y G S D A M A S 976 TACATTGGCAAACTTCGTATGGGTCGATGACTCCAATTGTGACAAATCAACAGTATCCTTTGACTAAGGGAATAA 325 V H W Q T S Y G S M T P I V T N Q Q Y P L T K G I 1051 TGCAAAATATTGTTCCGCCTAGTATGTCAATATACCAGCAAGATGGCTCTTCCTCACAGGGCACTCACCATTCAA 350 M Q N I V P P S M S I Y Q Q D G S S S Q G T H H S 1126 CACCTCAAGACAATGCAGGAATGGATGCTTCATTTCAAAGTAACTTGAAATAGCCAAACGCCACAGAGGGTGTAT 375 T P Q D N A G M D A S F Q S N L K * 1201 GA

6 GbAGL66 NtAGL66 TcAGL66 OsAGL66 PdAGL66 EgAGL66 PpAGL104 CmAGL66 GbAGL66 NtAGL66 TcAGL66 OsAGL66 PdAGL66 EgAGL66 PpAGL104 CmAGL66 (1) MGRVKLPIKKIENSTNRQVTFSKRRNGLIKKAYELSVLCDIEIALIMFSPSGRLSHFSGKNSRIEDVIARFVNLPEHERP (1) MGRVKLQIKKIENTTNRQVTFSKRRNGLIKKAYELSVLCDVDVALIMFSPSGRISTFSGNK-SIEDIMARYVNLPEHDRG (1) MGRVKLQIKRIENTTNRQVTFSKRRNGLIKKAYELSVLCDVDVAVMMFSPSGRVSLFSGNK-SIEEILGRYVNLPEHERG (1) MGRVKLPIKRIENTTNRQVTFSKRRNGLIKKAYELSVLCDIDVALLMFSPSGRLSHFSGRR-GVEDVILRYMNLSEHDRG (1) MGRVKLQIKRIENNTNRQVTFSKRRNGLIKKAYELSVLCDIDIALIMFSPSGRLSHFSGRQ-RIEDVLGRYINLPEHDRG (1) MGRVKLQIKRIENNTNRQVTFSKRRNGLIKKAYELSVLCDIDIALIMFSPSGRLSHFSGRR-RIEDVLARYISLPEHDRG (1) MGRVKLEIKRIENTTNRQVTFSKRRNGLIKKAYELSILCDIDIALIMFSPSGRLSHFSGKR-RIEDVFTRYINLPDQERE (1) MGRVKLQIKRIENTTNRQVTFSKRRNGLIKKAYELSILCDIDIALIMFSPSGRLSQFSGRR-RIEDVLARYINLPDHDRG (81) R-LVQNQEYLLRALKKLKYESDIANHLASPN---IVDSNVEELQMDIRRQRIQLEEAQQKLRSFKEDPLLITSIQDADQY (80) R--LHNQEHLQRAIAKLKCETDRTYQAASPAS---VDSQIEEFQQEILKYKTQLEDMDRRLRMYEGDLCEITTVCEAQYR (80) R--LRNKEFLLKALGKLRDETDQTCQAASPVS---TDSQLEEFQQEILKCKSRIVDMEKRLRVFEGDPFEITTLCQAEFH (80) E-AIQNREYLISMLQRLKRESDMATQLAN---PGALNEKIEEIQQEIYSSQQQLQITEDRLRMFEPDPAAFGTSSEVDGC (80) G-IIQNREYLIRTLKKLKCEGDMEAELVN---PGAVNFHVEELQQEIHMYQHQLQLSEERLSIFEPDPLGITSMDELESC (80) G-VIQNRDYLIRTLKMLKCESDMAAELVN---PGAVNSQVEELQQEIHRFQHQLQLSEERLRFFEPDPLSITSMDELESC (80) -HDLQNKEYLLRTLQQLRSENDIALQLANPTA--VSSE-IEELQQEIGGLQQQLQMAEEQIRIYEPDPLKITSMAEFESC (80) S-VVQNKEFLLGTLNNLKTENDIAQQLSNPTS--SNSN-VEELQQEVGTLRHELQLAEQQLRLFEPDFLSFTSNAEINSC K M GbAGL66 (157) ERTLEEALRRVRLRKQQLEHNQMAVAFN----D------ANLQFYIQTQN----GLPNGTDTSQNHLYNSWMPQGDPHTS NtAGL66 (155) EEILQETLKQVQARKHVLEETYSSPQRQTAPQPQMDFASQNVNVVNNVATSD------SIPNTSSNTFMDWVPRSQRDPH TcAGL66 (155) EQILEETLKQVRLRKQVLQEKYSSP----GLPP-----TTQVHLPPETADVN------GFVTGSSSSILEWIP--QRDPQ OsAGL66 (156) EKYLMELLTRVVERKNNLLSSHMAPFDA----T-----TAAMQGADGTQM--YVSQADGLATF-GGDAAMWGPDGGADPG PdAGL66 (156) EKFLMEALRRAGERKKYLLSNHLPPYDP----S-----ASSMQMYLPSQ------QEPMPNSF-GSEVVQWVPGTTPNPG EgAGL66 (156) EKFLMEALRRVAERKKYLLSNHLPSYDP----S-----TSSMQMYLQSQ------QEPMPNAF-GSEVVQWVPGTTPNPG PpAGL104 (156) EKSLMDTLTRVMQRKEYLLSNHLSSYD-----S------SGMPQVLPGSF ENEVAGWLS-SGGHNQ CmAGL66 (156) EKNLLDTLARITQRKKDLLSTHLSPYEP----P------NGIQIYLDQQD-----G--IPTSF-ESDVGNWLPENGQNNP GbAGL66 (223) VQNFMEHE-NSNAMLAMREAQCMAKCLQNGTVFPALQDATGMQLPNESASTQPYIPTSHMQFDYTLTDNNNNEHAEQADI NtAGL66 (229) VQILNFLDSNGLLPFRDEADQRMENMLPP-SLTQLHTPNVSAV VTDHLSPPNNRFE TcAGL66 (218) IQILNFLGSNGLLRPRDQSQPGAENILPPPQSTLLHGEEIN VDDQLSPRSG-LE OsAGL66 (224) HPMFSASDPLIYLR-----DHDVYDANSQVAGLHG----HAAAA DAWRQAYTCTEL PdAGL66 (220) NQIFIGSDLLMSLR-----DPGIYGPISSGTGLPMDPRVGSCHVSNQN DAWHQAYTSTEL EgAGL66 (220) HPIFIGSDPLMSLR-----DPGIYGPISSGAGLPVDTRLGSCHISNQN DTWHQAYTSTEL PpAGL104 (210) AQIYDASAPLDHQ--LRNLSSTLYDPFSQGTSSNADPSSMGECHVSVTNAS DGELPPWPQAYTNSSG CmAGL66 (218) NQICVASE-SSSIPQSGQ-----YPTTTVYDQVVSQAATTNINVGVGVGVG---VGGYDIANPNDDGFSPWHHNYTTTQL GbAGL66 (302) AAAFDYGSDAMASVHWQTSYGSMTPIVTNQQYPLTKGIMQNIVPPSMSIYQQDGSSSQGTHHSTPQDNAGMDASFQSNLK NtAGL66 (284) NDPPRPTSSFEGII DVNNAPWPPLYPTGNDPFPEAQPRERALLELFLSQLTPVNQDHL TcAGL66 (271) NDNIVQRPEFGQVV DVNLSPWTELYPTGDDSFPDAQPGGRALLELYLSQFTPSSISTMNQHQT OsAGL66 (271) LSTLIPTTPFPLMP-----HCLGPEDQYLSMEHGMVAAAQEPVEASTASCSYVPSDENSGTPVMAYDSN----PPPANIA PdAGL66 (275) LSALIPSAPFPLIQ-----HPMALTDLPPMVLPAEQAEAPT SCPHMPGDEH-GTCATTYETK----PPSVNAG EgAGL66 (275) LSALIPSAPFPLIQ-----HPMAPTDLSPMVPPAEQAKAQA SCPHMPGDEH-GTCATTYENN----PSSVNVG PpAGL104 (275) HPSTLMPSGLLP----QFQHTMVGSNMPEIMP-HEQVEIPVGS------PNVQAYN-EGAEYHENK VPQLNGH CmAGL66 (289) LSSFIPQTSFDV-----VKNEIGEPCMNTMIPQQQVDSISNGNQ MPPSDGSANYDNVK LSQLNVD K

7 25 AtAGL HbAGL30 HbAGL65 AtAGL65 AtAGL94 MIKC* GbAGL66 95 EgAGL PdAGL66 Type II 99 CaAG NaAG CsAGL3 ThAGL3 MIKC c 99 NoAGL6 100 CaAGL6 91 MhAGL6 72 AtAGL38 60 TgAGL45 AtAGL54 85 AtAGL AtAGL57 Type I AtAGL64 34 HaAGL AtAGL28 AcAGL73 Relative expression e d Stems Leaves Roots Male flowers a b c Female flowers d Young fruits

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