Nghiên cứu phân lập, giải trình tự và phân tích gen rpl16 ở một số mẫu giống nghệ (Curcuna longa L.) có hàm lượng curcuminoids khác nhau (Khóa luận tốt nghiệp)

Nghiên cứu phân lập, giải trình tự và phân tích gen rpl16 ở một số mẫu giống nghệ (Curcuna longa L.) có hàm lượng curcuminoids khác nhau (Khóa luận tốt nghiệp)Nghiên cứu phân lập, giải trình tự và phân tích gen rpl16 ở một số mẫu giống nghệ (Curcuna longa L.) có hàm lượng curcuminoids khác nhau (Khóa luận tốt nghiệp)Nghiên cứu phân lập, giải trình tự và phân tích gen rpl16 ở một số mẫu giống nghệ (Curcuna longa L.) có hàm lượng curcuminoids khác nhau (Khóa luận tốt nghiệp)Nghiên cứu phân lập, giải trình tự và phân tích gen rpl16 ở một số mẫu giống nghệ (Curcuna longa L.) có hàm lượng curcuminoids khác nhau (Khóa luận tốt nghiệp)Nghiên cứu phân lập, giải trình tự và phân tích gen rpl16 ở một số mẫu giống nghệ (Curcuna longa L.) có hàm lượng curcuminoids khác nhau (Khóa luận tốt nghiệp)Nghiên cứu phân lập, giải trình tự và phân tích gen rpl16 ở một số mẫu giống nghệ (Curcuna longa L.) có hàm lượng curcuminoids khác nhau (Khóa luận tốt nghiệp)

NGHIÊN PHÂN , RPL16 TRÌNH VÀ PHÂN TÍCH GEN (CURCUNA LONGA L.) CĨ HÀM : Chính quy Chun ngành Khoa : CNSH - CNTP : 2011 - 2015 NGHIÊN PHÂN RPL16 , TRÌNH VÀ PHÂN TÍCH GEN (CURCUNA LONGA L.) CĨ HÀM : Chính quy Chun ngành Khoa : CNSH - CNTP : K43 - CNSH : 2011 - 2015 êm i tích gen rpl Curcuna longa L i- Sinh viên ng ii 13 rpl16 19 19 19 22 23 25 rpl 34 iii H 22 24 25 26 rpl16 27 28 Hình 4.8 rpl 36 iv µg Micro gram µl Micro litre µM Micro Mole Bp CS DNA Deoxyribonucleic acid dNTPs deoxyribonucleotide triphosphates Kb Kilobase mM Mili Mole Na2EDTA Ethylene diamine tetra acetate sodium NST OD Optical density PCR polymerase chain reaction RNA ribonucleic acid TE Tris EDTA v .2 .3 .4 2.1 .8 11 11 13 13 13 14 14 14 vi 14 14 15 15 15 .17 .18 ® ExpinTM 20 21 22 .22 .22 23 rpl1624 24 27 .36 37 5.1 37 37 38 1 1.1 T Weir et al (2007) [32], Desmethoxycurcumin Bisdesmethoxycurcumin hóa (Shukla et al 2008)[28] (Kalpana et al 2004)[14] rpl Curcuna longa L.) có rpl Curcuna longa L 28 ng M Marker DNA (1 kb), Hình 4.6 N18 Qua Hình 4.6 ng 0,26 au tinh c rpl rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AAAATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAG GAAGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGC ATATTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGAT CCTTTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAA TCTAAAGTTTCGCGGGCGAATATTGACTCTTTC (264 bp) rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AAAATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAG GAAGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGC ATATTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGAT CCTTTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAA TCTAAAGTTTCGCGGGCGAATATTGACTCTTTC rpl 29 GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAGGA AGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGCATA TTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGATCCT TTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAATCT AAAGTTTCGCGGGCGAATATTGACTCTTTC (262 bp) rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAGGA AGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGCATA TTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGATCCT TTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAATCT AAAGTTTCGCGGGCGAATATTGACTCTTTC rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAGGA AGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGCATA TTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGATCCT TTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAATCT AAAGTTTCGCGGGCGAATNATTGACTCTTTC rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAGGA AGATGTATGTACGGGATATACAGATAGAATGTTTCTTTATGCATAT TTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGATCCTT TTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAATCTA AAGTTTCGCGGGCGAATATTGACTCTTTC 30 rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAGGA AGATGTATGTACGGGATATACAGATAGAATGTTTCTTTATGCATAT TTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGATCCTT TTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAATCTA AAGTTTCGCGGGCGAATATTGACTCTTTC Trìn rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AAAATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAG GAAGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGC ATATTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGAT CCTTTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAA TCTAAAGTTTCGCGGGCGATAATTGACTCTTTC rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AAAATAATCCAATAAGACATATTATCAATATTGAATTTGTTATATA GGAAGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATG CATATTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGA TCCTTTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTA ATCTAAAGTTTCGCGGGCGAAATATTGACTCTTTC rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AATAATCCATAAGACATTTATCAATATTGAATTTGTTATATAGGAA GATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGCATAT TTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGATCCTT 31 TTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAATCTA AAGTTTCGCGGGCGTTAAATTGACTCTTTC rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AAAATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAG GAAGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGC ATATTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGAT CCTTTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAA TCTAAAGTTTCGCGGGCGAATTTGGACTCTTTC rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AAAATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAG GAAGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGC ATATTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGAT CCTTTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAA TCTAAAGTTTCGCGGGCGAATATTGACTCTTTC rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AAAATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAG GAAGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGC ATATTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGAT CCTTTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAA TCTAAAGTTTCGCGGGCGAATATTGACTCTTTC Trì rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AAAATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAG 32 GAAGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGC ATATTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGAT CCTTTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAA TCTAAAGTTTCGCGGGCGAATATTGACTCTTTC rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AAAATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAG GAAGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGC ATATTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGAT CCTTTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAA TCTAAAGTTTCGCGGGCGAATATTGNACTCTTTC rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AAAATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAG GAAGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGC ATATTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGAT CCTTTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAA TCTAAAGTTTC rpl GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AAAATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAG GAAGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGC ATATTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGAT CCTTTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAA TCTAAAGTTTCGCGGGCGAATATTGGACTCTTTC rpl 33 GTTTCTTCTCATCCAGCTCCATGACATATTACAGATACACACGGAA AAAATAATCCAATAAGACATTTATCAATATTGAATTTGTTATATAG GAAGATGTATGTACGGGATATACAGATAGAATGTTTCTATTATGC ATATTTATGGATTATAGATAATGTTTATAATGTTTTTTTATAATGAT CCTTTTTTTGACCCTTCTCAAAGGATGCCAAAATATTGTAATGTAA TCTAAAGTTTCGCGGGCGAATATTGGACTCTTTC curc Hình 4.7 34 35 rpl rpl16 STT curcumin nucleotide gene N1 3,62 ± 0,02 GAAAAAAT 264 N2 2,05 ± 0,02 GAAAAAAT 264 N3 0,36 ± 0,01 GAAAAT 262 N4 0,38 ± 0,01 GAAAAT 262 N5 0,42 ± 0,02 GAAAAT 262 N6 0,48 ± 0,01 GAAAAT 262 N7 0,68 ± 0,01 GAAAAT 262 N8 1,98 ± 0,01 GAAAAAAT 264 N9 0,25 ± 0,02 GAAAAT 262 10 N10 6,35 ± 0,01 GAAAAAAT 264 11 N11 5,80 ± 0,01 GAAAAAAT 264 12 N12 5,75 ± 0,01 GAAAAAAT 264 13 N13 5,87 ± 0,01 GAAAAAAT 264 14 N14 4,27 ± 0,02 GAAAAAAT 264 15 N15 6,60 ± 0,01 GAAAAAAT 264 16 N16 7,25 ± 0,02 GAAAAAAT 264 17 N17 6,65 ± 0,02 GAAAAAAT 264 18 N18 8,32 ±0,02 GAAAAAAT 264 rpl16 - - 36 Hayakawa et al., n Hình 4.8 (A) (B) Hình 4.8 rpl (A) N18 (B) rpl16 8,32 %) N10, N11, N12, N13, N14, N15, N16, N17 N18 GAAAAAAT gene (0,25 37 5.1 1) cao (1,98 0,68%) 2) rpl 3) GAAAAAAT , hàm GAAAAT có kích 5.2 : rpl 38 I Côn - 2008, , trang 287 294 Curcuma longa Thanh Hóa , T (2009), o(1988 2008) Isoxaxolcurcumin Aggarwal B.B., Kumar A., Aggarwal M.S., Shishodia S (2005), Curcumin derived from turmeric (Curcuma longa) : a spice for all seasons, CRC Press LLC, Aoi K., Kaburagi K., Seki T., Tobata T., Sarak M and Kuroyanagi M (1986), Studies on the cultivation of turmeric (Curcuma longa L.): 39 Varietal differences in rhizome yield and curcuminoid content, Bulletin of National Institute of Hygienic Sciences, (104), pp 124-128 Apavatjrut P, Anuntalabhochai S, Sirirugsa P, Alisi C (1999) Molecular Curcuma (Zingiberaceae) species Ann Bot 84, pp 529 534 Chattopadhyay I., Biswas K., Bandyopadhyay U., and Banerjee R K (2004), Turmeric and curcumin: Biological actions and medicinal applications Curr Sci 87, pp 44 50, Chearwae W., Anuchapreedac S., Nandigama K., Ambudkar S.V., Limtrakul P (2004), Biochemical mechanism of modulation of human P-glycoprotein (ABCB1) by curcumin I, II, and III purified from Turmeric powder, Biochemical Pharmacology, Chen W-F., Deng S-L., Zhou B., Yang L., Liu Z-L., (2006), Curcumin and its analogues as potent inhibitors of low density lipoprotein oxidation: Hatom abstraction from the phenolic groups and possible involvement of the 4-hydroxy-3-methoxyphenyl groups, 40, pp 526-535 Cintra1 MMDF., Pinheiro J.B., and Sibov S.T (2005), Genetic divergence among Curcuma longa L.Accessions, Crop Breeding and Applied Biotechnology, 5, pp 410-417 Daniel V., Limson J.L., Dairam A., Watkins G.M., Daya S (2004), Through metal binding, curcumin protects against lead- and cadmiuminduced lipid peroxidation in rat brain homogenates and against leadinduced tissue damage in rat brain, Journal of Inorganic Biochemistry 98, pp 266-275 Hayakawa H, Kobayashi T, Minaniya Y, Ito K , Miyazaki A, Fukuda T, Yamamoto Y (2011a) Development of a molecular marker to identify a 40 candidate line of turmeric (Curcuma longa L.) with a high Curcumin content American Journal of Plant Sciences, (2): 15-26 10 Hayakawa H., Kobayashi T., Minamiya Y., Ito K., Miyazaki A., Fukuda T., Yamamoto Y (2010), Molercular Indentification of Turmeric ( Curcuma longa, Zingiberaceae) with High Curcumin Content, Journal of Janpanese Botany, 85(5), pp 263-269 11 Hayakawa H., Minaniya Y., Ito K., Yamamoto Y., Fukuda T (2011b), Difference of Curcumin content in Curcuma longa L (Zingiberaceae) caused by hybridization with other Curcumaspecies American Journal of Plant Sciences, (2), pp 111-119 12 Jayaprakasha GK., Negi PS., Anadharamakrishman C and Sakariah K.K (2001) Chemical composition of tumeric oil a hyproduct for from tumeric oleorsin inductry and Its inhibitory activity against different fungi Z Natuforsch., 56, pp 40-44 13 Joshi RK, Kuanar A, Mohanty S, Subudhi E, Nayak S (2010) Mining and characterization of EST derived microsatellites in Curcuma long a L Bioinformation 5(3): 128-131 14 Kalpana J., Warude P.C and Bhushan P (2004), Molecular markers in herbal drugtechnology Current Science, 87(2), pp 159-165 15 Kapoor LD (1990) Handbook of Ayurvedic Medicinal Plants, CRC Press, Boca Raton, Florida, 185 16 Lin, J K.; Lin-Shia, S Y., (2001) Mechanisms of cancer chemoprevention by curcumin, Proc Natl Sci Counc Repub China B 25, pp 59 66 17 Mahady G.B., Pendland S.L., Yun G And Lu Z.Z (2002) Tumeric (Curcuma longa L.) and curcumin inhibit the growth of Helico-bacter pylori, of group carcinogen Anticancer Res.,( 22), pp 4179-4181 41 18 Peter K.V., (2001), Hand book of herbs and spices, Woodhead Publishing Limited, Cambridge England 19 Pfeiffer E., Hohle S., Solyom, A., Metzler, M (2003), Studies on the stability of turmeric constituents J Food Eng 56, pp 257 259 20 Phan T., See P., Lee S., Chan S.Y (2001), Protective effects of curcumin against oxidative damage on skin cells in vitro: its implication for wound healing, J-Trauma 51, pp 927-931 21 Priyadarsini K.I., Mity D.K., Naik G.H., Kumar M.S., Unnikrishnan M.K., Satav J.G., Mohan H (2003), Role of phenolic O-H and methylene hydrogen on the free radical reactions and antioxidant activity of curcumin, Free Radical Biology and Medicine 35, pp 475-484 22 Ravindran P.N., Babu K.N and Sivaranan K (2007) Turmeric: The Genus Curcuma Medicinal and Aromatic Plants Industrial Profles CRC Press, Boca Raton 23 Reddy A.P., Lokesh B.R (1992), Studies on spice principles as antioxidants in the inhibition of lipid peroxidation of rat liver microsomes, Molecular and Cellular Biochemistry 111, pp 117-124 24 Runothayanun P., Wirachwong P (2005), Development of tetrahydrocurcuminoid liposomes as an ingredient for cosmetic products, 7th ASCS conference Bangkok, Thailand 25 Sasaki Y., Fushimi H., Cao H., Cai S.Q and Komatsu K (2002), Sequence Analysis of Chinese and Japanese Curcuma Drugs on the 18S rRNA Gene and trnK Gene and the Application of Amplification- Refractory Mutation System Analysis for Their Authentication Biological & Pharmaceutical Bulletin, 25, (12), pp 1593-1599 26 Selvam C., Jachak SM., Thilagavathi R., Chakraborti AK (2005), Design, synthesis, biological evaluation and molecular docking of curcumin 42 analogues as antioxidant, cyclooxygenase inhibitory and anti- inflammatory agents , Bioorganic & Medicinal Chemistry Letters 15, pp 1793-1797 27 Shen L., Ji H-F.(2007), Theoretical study on physicochemical properties of curcumin, Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 67, pp 619-623 28 Shukla P.K., Khanna V.K., Ali M.M., Khan M.Y., Srimal R.C (2008) Anti-ischemic effect of curcumin in rat brain, Neurochern Res 6/2008; 33(6), pp 1036-43 29 Sigrist M.S., Pinheiro J.B., Filho J.A and Zucchi M.I (2011) Genetic diversity of turmeric germplasm (Curcuma longa; Zingiberaceae) identifed by microsatellite markers Genetics and Molecular Research, 10 (1), pp 419-428 30 Srivatsava S and Nidhi M.(2009) Genetic Markers A Cutting Edge Technology in Herbal Drug Research Journal of Chemical and Pharmaceutical Research 1(1), pp 1-18 31 Tharachand C., Immanuel S.C and Mythili M.N (2012) Molecular markers in characterization of medicinal plants: An overview Research in Plant Biology, 2(2), pp 01-12 32 Weir N.M.,Selvendiran K.,Kutala V.K.,Tong L.,Vishwanath S.,Rajaram M.,Tridandapani S.,AnantS and Kuppusamy P (2007) CurcuminInducesG2/MArrestand Apoptosis in Cisplatin - Resistant Human Ovarian Cancer CellsbyModulating Akt andp38MAPK, CancerBiol- ogyandTherapy, (2), pp 178-184 33 Wright J.S (2002) Predicting the antioxidant activity of curcumin and curcuminoids J Mol Struct (Theochem.) , pp 207 217 ...NGHIÊN PHÂN RPL16 , TRÌNH VÀ PHÂN TÍCH GEN (CURCUNA LONGA L.) CĨ HÀM : Chính quy Chun ngành Khoa : CNSH - CNTP : K43 - CNSH : 2011 - 2015 êm i tích gen rpl Curcuna longa L i- Sinh... 2004)[14] rpl Curcuna longa L.) có rpl Curcuna longa L 3 1.2.2 rpl16 rpl 1.3.1 rpl ông 1.3.2 - rpl - - vàng Curcuma longa cs 2004)[1] cs (2004)[1], Á H có 16 lồi (Curcuma longa L aromatica S xanhthorrhiza... loài C longa Hayakawa et al.(2011b)[11] phân tích gene matK c ETS mã hóa cho riboxom C longa hác chi Curcuma Curcuma longa trung bình cao Aoi et al.(1986)[2] C longa Cintra et al.(2005)[7] ác nghiên