Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.

ÖKTEM, HÜSEYİN AVNİ; Baloglu, Mehmet Cengiz; YÜCEL, Ayşe Meral; ZAKHAROV, Florence NEGRE

Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.

Mehmet Cengiz BALOĞLU, (Orta Doğu Teknik Üniversitesi, Biyolojik Bilimler Bölümü, Ankara, Türkiye)

Florence NEGRE ZAKHAROV, (Department, of Plant Sciences, Univercity of California at Davis, One Shields Avenue, Davis, CA 95616 USA)

Hüseyin Avni ÖKTEM, (Orta Doğu Teknik Üniversitesi, Biyolojik Bilimler Bölümü, Ankara, Türkiye)

Ayşe Meral YÜCEL (Orta Doğu Teknik Üniversitesi, Biyolojik Bilimler Bölümü, Ankara, Türkiye)

Turkish Journal of Biology

3 0

Yıl: 2011 Cilt: 35 Sayı: 4 Sayfa Aralığı: 387 - 397 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.

Öz:

GTP-bağlayıcı protein ailesinin yüksek derecede korunmuş üyesi olan Ran bağlayıcı proteinleri (RanBPs) stoplazma ve çekirdek arasındaki çekirdek proteinlerin ihracatına katılmaktadırlar. Bu çalışmada, kavundan (Cucumis melo L.) RACE metodu kullanılarak CmRanBP geni izole edilmiştir. 778 baz çift i uzunluğunda ve 197 amino asiti (22,2 kDa protein) kodlayan kavun RanBP cDNA’sı karakterize edilmiştir (GenBank erişim no: EU853459). CmRanBP genine ait tahmini aminoasit sekansının Vitis vinifera, Populus trichocarpa ve Ricinus communis bitkilerine ait sırasıyla VvRanBP, PtRanBP, and RcRanBP genleri ile % 70 oranında benzerlik gösterdiği bulunmuştur. Çeşitli bitki türlerinde RanBD (Ran bağlayıcı domain) içinde RanBP gen ailesinin bütün üyelerinde yüksek oranda korunumlu beş motif ve bir tane Ran bağlayıcı motif bulunmuştur. Yüksek sıcaklık stresi altında farklı dokularda CmRanBP gen ifade profi lleri de yarı kantitatif RT-PCR ile incelendi. Kontrol ortamı olarak 25 °C’de CmRanBP geni kök, yaprak ve gövdelerde benzer bir şekilde ifade edilmiştir. Ancak, sıcaklık 38 °C ve 40 °C’ ye yükseltildiği zaman, kök, yaprak ve gövde dokularında CmRanBP gen ifade düzeyi anlamlı (P < 0,05) şekilde artmıştır. Biz burada ilk kez CmRanBP gen ifadesinin ısı stres yanıtları ile ilişkili olduğunu göstermekteyiz.

Anahtar Kelime: DNA klonlama tamamlayıcı DNA Cucumis melo kavun sıcaklık stresi RNA izolasyonu gen ifadesi bağlayıcı proteinler filogenetik bitkiler

Konular: Biyoloji

Kavunda (Cucumis melo L.) Ran bağlayıcı proteini (RanBP) kodlayan genin moleküler klonlanması, karakterizasyonu ve ifade analizi

Öz:

Ran binding proteins (RanBPs) are highly conserved members of the GTP-binding protein family that are involved in nuclear protein export between the nucleus and the cytoplasm. In this study, a CmRanBP gene from a melon was isolated (Cucumis melo L.) using the RACE (rapid amplifi cation of cDNA ends) method. Th e 778 basepair long melon, with a RanBP cDNA encoding consisting of 197 amino acids (22.2 kDa protein), was characterized (GenBank accession no: EU853459). Th e predicted amino acid sequence of CmRanBP was found to be 70% identical to VvRanBP, PtRanBP, and RcRanBP from Vitis vinifera, Populus trichocarpa, and Ricinus communis, respectively. Within the RanBD (Ran binding domain), 5 highly conserved motifs and 1 Ran binding motif were found in all members of the RanBP gene family from various plant species. Expression profi les of the CmRanBP gene in diff erent tissues under high temperature stress were also investigated by semiquantitative RT-PCR. Th e CmRanBP gene was expressed in a similar manner in the roots, leaves, and stems at 25 °C as a control environment. However, when the temperature was raised to 38 °C and 40 °C, expression levels of the CmRanBP gene were signifi cantly (P < 0.05) increased in the root, leaf, and stem tissues. We show here for the fi rst time that the CmRanBP gene expression was correlated with heat stress responses.

Anahtar Kelime: heat stress RNA isolation gene expression binding proteins phylogenetics plants DNA cloning complementary DNA Cucumis melo melons

Konular: Biyoloji

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

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APA	Baloglu M, ZAKHAROV F, ÖKTEM H, YÜCEL A (2011). Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.. , 387 - 397.
Chicago	Baloglu Mehmet Cengiz,ZAKHAROV Florence NEGRE,ÖKTEM HÜSEYİN AVNİ,YÜCEL Ayşe Meral Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.. (2011): 387 - 397.
MLA	Baloglu Mehmet Cengiz,ZAKHAROV Florence NEGRE,ÖKTEM HÜSEYİN AVNİ,YÜCEL Ayşe Meral Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.. , 2011, ss.387 - 397.
AMA	Baloglu M,ZAKHAROV F,ÖKTEM H,YÜCEL A Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.. . 2011; 387 - 397.
Vancouver	Baloglu M,ZAKHAROV F,ÖKTEM H,YÜCEL A Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.. . 2011; 387 - 397.
IEEE	Baloglu M,ZAKHAROV F,ÖKTEM H,YÜCEL A "Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.." , ss.387 - 397, 2011.
ISNAD	Baloglu, Mehmet Cengiz vd. "Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.". (2011), 387-397.

APA	Baloglu M, ZAKHAROV F, ÖKTEM H, YÜCEL A (2011). Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.. Turkish Journal of Biology, 35(4), 387 - 397.
Chicago	Baloglu Mehmet Cengiz,ZAKHAROV Florence NEGRE,ÖKTEM HÜSEYİN AVNİ,YÜCEL Ayşe Meral Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.. Turkish Journal of Biology 35, no.4 (2011): 387 - 397.
MLA	Baloglu Mehmet Cengiz,ZAKHAROV Florence NEGRE,ÖKTEM HÜSEYİN AVNİ,YÜCEL Ayşe Meral Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.. Turkish Journal of Biology, vol.35, no.4, 2011, ss.387 - 397.
AMA	Baloglu M,ZAKHAROV F,ÖKTEM H,YÜCEL A Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.. Turkish Journal of Biology. 2011; 35(4): 387 - 397.
Vancouver	Baloglu M,ZAKHAROV F,ÖKTEM H,YÜCEL A Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.. Turkish Journal of Biology. 2011; 35(4): 387 - 397.
IEEE	Baloglu M,ZAKHAROV F,ÖKTEM H,YÜCEL A "Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.." Turkish Journal of Biology, 35, ss.387 - 397, 2011.
ISNAD	Baloglu, Mehmet Cengiz vd. "Molecular cloning, characterization, and expression analysis of a gene encoding a Ran binding protein (RanBP) in Cucumis melo L.". Turkish Journal of Biology 35/4 (2011), 387-397.