scholarly journals STUDI BIOINFORMASI URUTAN ASAM AMINO DAN STRUKTUR 3D PROTEIN ALDOLASE KELAS II (AldII) DARI Uncultured Acidilobus sp.

2021 ◽  
Vol 6 (2) ◽  
pp. 117
Author(s):  
Nishia Waya Meray ◽  
Suharti Suharti ◽  
Akhmaloka Akhmaloka
Keyword(s):  
Ab Initio ◽  
Open Reading Frame ◽  
Reading Frame ◽  
Β Sheet

Pada penelitian sebelumnya fragmen gen 1,9 kb telah berhasil diisolasi dari Kawah Domas, Jawa Barat melalui pendekatan metagenom. Fragmen tersebut diketahui mengandung daerah Open Reading Frame (ORF) utuh dari gen pengkode aldolase kelas II dari uncultured Acidilobus sp. yang kemudian disebut sebagai aldII. Fragmen gen aldII tersebut berhasil diekspresikan menjadi protein termostabil aldolase kelas II yang kemudian disebut sebagai AldII. Penelitian ini bertujuan untuk melakukan studi bioinformasi terhadap protein AldII tersebut. Protein AldII kemudian diketahui memiliki massa molekul ~21,2 kDa dengan rumus molekul C940H1539N261O281S8. Total residu bermuatan negatif (Asp + Glu) sebanyak 22 residu, sedangkan total residu bermuatan positif (Arg + Lys) adalah 18 residu. Nilai pI teoritis AldII sebesar 5,86. Hasil perhitungan indeks kestabilan protein ini adalah 36,61 dan diklasifikasikan sebagai protein yang stabil. Lewat penjajaran dengan homologi terdekat, ditemukan daerah lestari yang dapat menunjukan residu yang mungkin berperan dalam pengikatan logam dan sisi aktif. Prediksi struktur 3D dilakukan secara ab initio, menunjukan adanya 6 struktur β-sheet dan 6 struktur α-heliks. Dengan demikian dapat disimpulkan bahwa protein AldII dari uncultured Acidilobus sp. diduga memiliki aktivitas enzimatik.

scholarly journals The cDNA for rat selenoprotein P contains 10 TGA codons in the open reading frame

1991 ◽  
Vol 266 (16) ◽  
pp. 10050-10053
Author(s):  
K.E. Hill ◽  
R.S. Lloyd ◽  
J.G. Yang ◽  
R. Read ◽  
R.F. Burk
Keyword(s):  
Open Reading Frame ◽  
Selenoprotein P ◽  
Reading Frame

scholarly journals The Cloning and Molecular Analysis of pawn-B in Paramecium tetraurelia

Genetics ◽  
2000 ◽  
Vol 155 (3) ◽  
pp. 1105-1117 ◽  
Author(s):  
W John Haynes ◽  
Kit-Yin Ling ◽  
Robin R Preston ◽  
Yoshiro Saimi ◽  
Ching Kung
Keyword(s):  
Genomic Library ◽  
Open Reading Frame ◽  
Paramecium Tetraurelia ◽  
Wild Type ◽  
Rt Pcr ◽  
Reading Frame ◽  
Close Proximity ◽  
In The Wild ◽  
Dna Fragment ◽  
Alternative Sites

Abstract Pawn mutants of Paramecium tetraurelia lack a depolarization-activated Ca2+ current and do not swim backward. Using the method of microinjection and sorting a genomic library, we have cloned a DNA fragment that complements pawn-B (pwB/pwB). The minimal complementing fragment is a 798-bp open reading frame (ORF) that restores the Ca2+ current and the backward swimming when expressed. This ORF contains a 29-bp intron and is transcribed and translated. The translated product has two putative transmembrane domains but no clear matches in current databases. Mutations in the available pwB alleles were found within this ORF. The d4-95 and d4-96 alleles are single base substitutions, while d4-662 (previously pawn-D) harbors a 44-bp insertion that matches an internal eliminated sequence (IES) found in the wild-type germline DNA except for a single C-to-T transition. Northern hybridizations and RT-PCR indicate that d4-662 transcripts are rapidly degraded or not produced. A second 155-bp IES in the wild-type germline ORF excises at two alternative sites spanning three asparagine codons. The pwB ORF appears to be separated from a 5′ neighboring ORF by only 36 bp. The close proximity of the two ORFs and the location of the pwB protein as indicated by GFP-fusion constructs are discussed.

scholarly journals GIT1, a Gene Encoding a Novel Transporter for Glycerophosphoinositol in Saccharomyces cerevisiae

Genetics ◽  
1998 ◽  
Vol 149 (4) ◽  
pp. 1707-1715 ◽  
Author(s):  
J L Patton-Vogt ◽  
S A Henry
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Regulatory Gene ◽  
Sugar Transport ◽  
Open Reading Frame ◽  
Gene Encoding ◽  
Reading Frame ◽  
Membrane Spanning ◽  
Membrane Spanning Domains ◽  
Uptake And Metabolism ◽  
Cells Cultured

Abstract Phosphatidylinositol catabolism in Saccharomyces cerevisiae cells cultured in media containing inositol results in the release of glycerophosphoinositol (GroPIns) into the medium. As the extracellular concentration of inositol decreases with growth, the released GroPIns is transported back into the cell. Exploiting the ability of the inositol auxotroph, ino1, to use exogenous GroPIns as an inositol source, we have isolated mutants (Git−) defective in the uptake and metabolism of GroPIns. One mutant was found to be affected in the gene encoding the transcription factor, SPT7. Mutants of the positive regulatory gene INO2, but not of its partner, INO4, also have the Git− phenotype. Another mutant was complemented by a single open reading frame (ORF) termed GIT1 (glycerophosphoinositol). This ORF consists of 1556 bp predicted to encode a polypeptide of 518 amino acids and 57.3 kD. The predicted Git1p has similarity to a variety of S. cerevisiae transporters, including a phosphate transporter (Pho84p), and both inositol transporters (Itr1p and Itr2p). Furthermore, Git1p contains a sugar transport motif and 12 potential membrane-spanning domains. Transport assays performed on a git1 mutant together with the above evidence indicate that the GIT1 gene encodes a permease involved in the uptake of GroPIns.

scholarly journals Antigenic Characterization of ORF2 and ORF3 Proteins of Hepatitis E Virus (HEV)

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1385
Author(s):  
Giulia Pezzoni ◽  
Lidia Stercoli ◽  
Eleonora Pegoiani ◽  
Emiliana Brocchi
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Recombinant Antigen ◽  
Open Reading Frame ◽  
Reading Frame ◽  
E Coli ◽  
Antigenic Characterization ◽  
Antigenic Properties ◽  
Open Reading Frame 2

To evaluate the antigenic properties of Hepatitis E Virus (HEV) Open Reading Frame 2 and 3 (ORF2 and ORF3) codified proteins, we expressed different portions of ORF2 and the entire ORF3 in E. coli, a truncated ORF2, was also expressed in baculovirus. A panel of 37 monoclonal antibodies (MAbs) was raised against ORF2 (1–660 amino acids) and MAbs were mapped and characterized using the ORF2 expressed portions. Selected HEV positive and negative swine sera were used to evaluate ORF2 and ORF3 antigens’ immunogenicity. The MAbs were clustered in six groups identifying six antigenic regions along the ORF2. Only MAbs binding to the sixth ORF2 antigenic region (394–608 aa) were found to compete with HEV positive sera and efficiently catch the recombinant antigen expressed in baculovirus. The ORF2 portion from 394–608 aa demonstrated to include most immunogenic epitopes with 85% of HEV positive swine sera reacting against the region from 461–544 aa. Only 5% of the selected HEV sera reacted against the ORF3 antigen.

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