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1.
Genet. mol. res. (Online) ; 2(4): 376-382, Dec. 2003.
Article in English | LILACS | ID: lil-417592

ABSTRACT

Pyrophosphate-dependent phosphofructokinase (PPi-PFK) has been detected in several types of plant cells, but the gene has not been reported in sugar cane. Using Citrus paradisi PPi-PFK gene (AF095520 and AF095521) sequences to search the sugar cane EST database, we have identified both the alpha and beta subunits of this enzyme. The deduced amino acid sequences showed 76 and 80 similarity with the corresponding alpha and beta subunits of C. paradisi. A high degree of similarity was also observed among the PFK b subunits when the alignment of the sugar cane sequences was compared to those of Ricinus communis and Solanum tuberosum. It appears that alpha and beta are two distinct subunits; they were found at different concentrations in several sugar cane tissues. It remains to be determined if the different gene expression levels have some physiological importance and how they affect sucrose synthesis, export, and storage in vacuoles. A comparison between the amino acid sequences of b PFKs from a variety of organisms allowed us to identify the two critical Asp residues typical of this enzyme's activity site and the other binding sites; these residues are tightly conserved in all members of this protein family. Apparently, there are catalytic residues on the b subunit of the pyrophosphate-dependent enzyme


Subject(s)
Phosphotransferases/genetics , Pyrophosphatases/metabolism , Saccharum/enzymology , Amino Acid Sequence , DNA, Complementary/analysis , Phosphotransferases/metabolism , Molecular Sequence Data , Saccharum/genetics
2.
Genet. mol. res. (Online) ; 1(4): 327-336, Dec. 2002.
Article in English | LILACS | ID: lil-417633

ABSTRACT

The effects of breed and of recombinant bovine somatotropin (rbST) treatment on growth hormone gene expression were studied in young bulls. The experiment was completely randomized in a [2 x 2]-factorial arrangement, using two levels of rbST (0 or 250 mg/animal/14 days), and two breed groups (Nelore and Simmental x Nelore crossbred). A cDNA encoding Bos indicus growth hormone was cloned and sequenced for use as a probe in Northern and dot blot analyses. Compared to the Bos taurus structural gene, the Bos indicus cDNA was found to begin 21 bases downstream from the transcription initiation site and had only two discrepancies (C to T at position 144-His and T to C at position 354-Phe), without changes in the polypeptide sequence. However, two amino acid substitutions were found for Bubalus spp., which belong to the same tribe. The rbST treatment did not change any of the characteristics evaluated (body and pituitary gland weights, growth hormone mRNA expression level). Crossbred animals had significantly higher body weight and heavier pituitaries than Nelore cattle. Pituitary weight was proportional to body weight in both breed groups. Growth hormone mRNA expression in the pituitary was similar (P>0.075) for both breed and hormonal treatment groups, but was 31.9 higher in the pure Nelore group, suggesting that growth hormone gene transcription regulation differs among these breeds


Subject(s)
Humans , Male , Cattle/growth & development , Gene Expression/drug effects , Pituitary Gland/drug effects , Growth Hormone/pharmacology , Cattle/genetics , DNA, Complementary/analysis , DNA, Complementary/genetics , Gene Expression/genetics , Pituitary Gland , Growth Hormone/genetics , Body Weight/drug effects , Body Weight/genetics , RNA, Messenger/drug effects , RNA, Messenger/genetics , Sequence Analysis, DNA
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