scholarly journals Cloning and Characterization of Partial Chlorophyll a Oxygenase (CAO) Gene Involved in Soybean Shade Tolerance Mechanism

2015 ◽  
Vol 2 (2) ◽  
pp. 1-4 ◽  
Author(s):  
Nurul Khumaida ◽  
Kisman Kisman ◽  
Didy Sopandie
Keyword(s):  
Chlorophyll A ◽  
Shade Tolerance ◽  
Leukemia Virus ◽  
Blot Hybridization ◽  
Rnase H ◽  
Moloney Murine Leukemia Virus ◽  
Northern Blot Hybridization ◽  
Plant Tolerance ◽  
Tolerance Mechanism ◽  
Soybean Plants

Plant tolerance mechanism under shade stress is indicated by high chlorophyll-b content, which is synthesized from chlorophyll a by chlorophyll a-oxygenase (CAO) gene. The study was aimed to clone and characterize the partial of CAO gene involved in shade tolerance mechanism in soybean.  Total RNA was isolated from the second trifoliate leaves of soybean plants and the first strand cDNA was prepared using Reverse Transcriptase Moloney Murine Leukemia Virus (RT-M-MLV) (RNase H-). Northern blot hybridization was used to analyze expression of partial CAO gene. The result showed that one partial gene, CAO 3-4 (1,052 bp), comprised of 292 adenine (a), 241 cytosine (c), 276 guanine (g) and 242 thymine (t), therefore the number of amino acid deduction was 338.  The expression of CAO 3-4 partial gene was higher in shade tolerant genotypes than in shade sensitive genotypes, both under 50% shade and dark. These results demonstrated that the CAO gene is involved in shade tolerance mechanism of soybean.

scholarly journals Preparation and characterization of the RNase H domain of Moloney murine leukemia virus reverse transcriptase

2015 ◽  
Vol 113 ◽  
pp. 44-50 ◽  
Author(s):  
Kosaku Nishimura ◽  
Kanta Yokokawa ◽  
Tetsuro Hisayoshi ◽  
Kosuke Fukatsu ◽  
Ikumi Kuze ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Murine Leukemia Virus ◽  
Leukemia Virus ◽  
Rnase H ◽  
Moloney Murine Leukemia Virus ◽  
Murine Leukemia

scholarly journals Small Dumbbell Oligonucleotides Inhibitors of RNase H Activity of MOMULV Reverse Transcriptase

2011 ◽  
Vol 8 (2) ◽  
pp. 629-634
Author(s):  
Ajay Kumar
Keyword(s):  
Reverse Transcriptase ◽  
Murine Leukemia Virus ◽  
Dissociation Constants ◽  
Leukemia Virus ◽  
Rnase H ◽  
Moloney Murine Leukemia Virus ◽  
Mobility Shift ◽  
Hiv 1 ◽  
Murine Leukemia

The small dumbbell oligonucleotides containing loops of phosphodiester (OL-1), two trimethylene, C3moieties in each loop (OL-2) and phosphorothioate (OL-3) linkages were synthesized. Incubation of OL-1 and OL-2 with S-1 nuclease generated break down products whereas incubation of OL-3 did not result in significant cleavage. Their binding to moloney murine leukemia virus reverse transcriptase was evaluated by PAGE band mobility shift assays. The OL-3 bound more strongly to the reverse transcriptase than OL-1 and OL-2. The dissociation constants evaluated using PAGE band mobility shift assays were of the order of 10-7. Investigation of inhibition of RNase H activity of reverse transcriptase showed that the OL-3 is a better inhibitor of the retroviral RNase H activity than both OL-1 and OL-2. Thus OL-3 may be used as RNase H inhibitor. Our studies demonstrated that this particularly designed oligonucleotide (OL-3) displays an IC50of 25 nM in its inhibition on the reverse transcriptase RNase H activity, a magnitude lower than that of first nucleotide reverse transcriptase of HIV-1, tenofovir, introduced by Gilead Science in the market.

Identification and chromosomal localization of a transcriptionally active retrotransposon of Ty3-gypsy type in rice

Genome ◽  
2000 ◽  
Vol 43 (2) ◽  
pp. 404-408 ◽  
Author(s):  
Zi-Yin Li ◽  
Shou-Yi Chen ◽  
Xian-Wu Zheng ◽  
Li-Huang Zhu
Keyword(s):  
Oryza Sativa ◽  
Copy Number ◽  
Oryza Sativa L ◽  
Blot Hybridization ◽  
Rice Genome ◽  
Rnase H ◽  
Northern Blot Hybridization ◽  
Japonica Rice ◽  
Terminal Part ◽  
Dot Blot

A DNA fragment representing a transcriptionally active retrotransposon of Ty3-gypsy type was isolated and characterized from rice (Oryza sativa L.). The fragment (named RIRE9) includes the coding sequences for the C-terminal part of the RNase H domain and the N-terminal part of the integrase domain in the polyprotein region. Northern blot hybridization indicated that this element was expressed in rice leaves and stems, suggesting that it is potentially active to transpose under normal growth conditions. Using dot-blot hybridization, the copy number of RIRE9 was estimated to be about 1600 copies per haploid rice genome. Five homologous copies of RIRE9 were assigned to five distinct positions of four chromosomes by restriction fragment length polymorphism (RFLP) mapping approach using an indica-japonica rice doubled-haploid (DH) population and its molecular linkage map. Key words: Oryza sativa L., Ty3-gypsy-like retrotransposon, copy number, chromosomal location.

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