• Title/Summary/Keyword: pMJ32E

Search Result 4, Processing Time 0.027 seconds

Construction of a Novel Shuttle Vector for Tetragenococcus species based on a Cryptic Plasmid from Tetragenococcus halophilus

  • Min Jae Kim;Tae Jin Kim;Yun Ji Kang;Ji Yeon Yoo;Jeong Hwan Kim
    • Journal of Microbiology and Biotechnology
    • /
    • v.33 no.2
    • /
    • pp.211-218
    • /
    • 2023
  • A cryptic plasmid (pTH32) was characterized from Tetragenococcus halophilus 32, an isolate from jeotgal, Korean traditional fermented seafood. pTH32 is 3,198 bp in size with G+C content of 35.84%, and contains 4 open reading frames (ORFs). orf1 and orf2 are 456 bp and 273 bp in size, respectively, and their translation products showed 65.16% and 69.35% similarities with RepB family plasmid replication initiators, respectively, suggesting the rolling-circle replication (RCR) mode of pTH32. orf3 and orf4 encodes putative hypothetical protein of 186 and 76 amino acids, respectively. A novel Tetragenococcus-Escherichia coli shuttle vector, pMJ32E (7.3 kb, Emr), was constructed by ligation of pTH32 with pBluescript II KS(+) and an erythromycin resistance gene (ErmC). pMJ32E successfully replicated in Enterococcus faecalis 29212 and T. halophilus 31 but not in other LAB species. A pepA gene, encoding aminopeptidase A (PepA) from T. halophilus CY54, was successfully expressed in T. halophilus 31 using pMJ32E. The transformant (TF) showed higher PepA activity (49.8 U/mg protein) than T. halophilus 31 cell (control). When T. halophilus 31 TF was subculturd in MRS broth without antibiotic at 48 h intervals, 53.8% of cells retained pMJ32E after 96 h, and only 2.4% of cells retained pMJ32E after 14 days, supporting the RCR mode of pTH32. pMJ32E could be useful for the genetic engineering of Tetragenococcus and Enterococcus species.

Duckweed as a Protein Source for Fine-Wool Merino Sheep: Its Edibility and Effects on Wool Yield and Characteristics

  • Damry, J.V. Nolan;Bell, R.E.;Thomson, E.S.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.14 no.4
    • /
    • pp.507-514
    • /
    • 2001
  • Two experiments were carried out to investigate whether duckweed is useful as a dietary protein source for fine-wool Merino sheep and to evaluate its effects on wool yield and characteristics. In Experiment 1, the sheep were given one of three maintenance diets consisting of oaten chaff (520-700 g/d) supplemented with 16-32 g crude protein/d in the form of fresh (1 kg/d) or sun-dried (50-100 g/d) duckweed. Each ration was estimated to provide 5.4 MJ (1.3 Mcal)/d of metabolisable energy (ME). The sheep readily ingested the fresh or dried duckweed. None of the wool measures (yield, rate of fibre elongation, fibre diameter) differed (p>0.05) between dietary treatments. In Experiment 2, oaten-chaff-based diets (800 g/d) supplying 6.5-7.2 MJ (1.6-1.7 Mcal)/d of ME were supplemented with iso-nitrogenous amounts (4-5 g N) either of urea (8 g), cottonseed meal (60 g) or dried duckweed (100 g). In this experiment, the rate of wool fibre elongation, thought to be related to intestinal amino acid absorption, was lower (p<0.05) for sheep given the oaten chaff/urea diet than for those given either oaten chaff/cottonseed meal or oaten chaff/duckweed for which the rates did not differ (p>0.05). Fibre diameter, which ranged from 16.0-16.7 mm, did not differ (p>0.05) between diets, but tended to be lower on the oaten chaff/urea diet so that volume of wool produced was also significantly lower (p<0.05) on this diet than on the diets containing duckweed or cottonseed meal. Rumen ammonia concentrations at 4.5 and 7.5 h after feeding were higher (p<0.05) for sheep given the oaten chaff/urea diet than for those given the other two diets. A comparison of the rumen ammonia concentrations, wool growth rate and predicted flows of amino acids from the rumen of sheep supplemented with duckweed rather than cottonseed meal suggested that duckweed is a valuable source of 'escape protein' for ruminants.

Variations in the Production, Qualitative Characteristics and Coagulation Parameters of the Milk of the Riverine Buffalo Determined by the Energy/Protein Content of the Diet

  • Bartocci, S.;Terramoccia, S.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.23 no.9
    • /
    • pp.1166-1173
    • /
    • 2010
  • Sixteen Mediterranean pluriparous buffaloes were subdivided into two uniform groups of eight animals. The average weight of the two groups at the start of the trial was 671.2 and 656.7 kg. The number of days from calving were 33.4 and 33.3, and the average milk production was 12.73 and 12.33 kg/d. The trial lasted for 114 days, and was divided into two sub-periods of 58 and 56 days. The two diets, administered ad libitum, had the same forage/concentrate ratio (53/47) but in their formulation the percentage of the two forages varied. Diet 1: alfalfa hay = 10%, maize silage = 43%, concentrate 1 = 47% (6.63 MJ/kg DM of net energy; 179.5 g/kg DM of crude protein). Diet 2: alfalfa hay = 20%, maize silage = 33%, concentrate 2 = 47%, (5.99 MJ/kg DM of net energy; 155.4 g/kg DM of crude protein). For the overall trial period (33-146 days in milk), the intake of dry matter was 17.23 kg/d for Group 1 and 17.29 kg/d for Group 2 and corresponded to 2.50 and 2.58% (p<0.01) of live weight. There was no significant difference between the average weight (689.7 and 669.4 kg) and the body condition score (6.49 and 6.42) of the two groups of buffaloes. Group 1 produced a greater quantity of milk (11.89 vs. 10.90 kg/d, p<0.10) of better quality both for its higher fat content (82.32 vs. 77.29 g/kg, p<0.10) and its protein content (47.36 and 46.38 g/kg). The milk produced by the buffaloes receiving Diet 1 had a better clotting ability, lower values of r (15.98 and 16.42 min) and K20 (1.66 and 1.75 min) and a higher value of A30 (54.45 and 52.73 mm). Taking into consideration the two sub-periods, milk production was significantly different only in the first sub-period (33-90 DIM), in favour of Group 1 (13.08 vs. 11.56 kg/d, p<0.05), while the positive effect of Diet 1 was cancelled out (10.71 and 10.24 kg/d) in the second part of the trial (91-146 DIM).

Comparison of In vitro Gas Production, Metabolizable Energy, Organic Matter Digestibility and Microbial Protein Production of Some Legume Hays

  • Karabulut, Ali;Canbolat, Onder;Kalkan, Hatice;Gurbuzol, Fatmagul;Sucu, Ekin;Filya, Ismail
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.20 no.4
    • /
    • pp.517-522
    • /
    • 2007
  • The aim of this study was to compare in vitro gas production kinetics, metabolizable energy (ME), organic matter digestibility (OMD) and microbial protein (MP) production of widely used legume hays in ruminant nutrition in Turkey. Gas production were determined at 0, 3, 6, 12, 24, 48, 72 and 96 h and their kinetics were described using the equation p = a+b ($1-e^{-ct}$). There were significant differences among legume hays in terms of chemical composition. The crude protein content of legume hays ranged from 11.7 to 18.6% of dry matter (DM); crude fat from 2.1 to 3.5% DM; neutral detergent fiber from 35.6 to 52.0% DM; acid detergent fiber from 32.0 to 35.5% DM and acid detergent lignin 1.7 to 11.0% DM. Total gas production after 96 h incubation ranged between 61.67 and 76.00 ml/0.200 g of substrate. At 24, 72 and 96 h incubation the total gas production for common vetch were significantly (p<0.01) higher than those of the other legume hays. The ME, OMD and MP of legume hays ranged from 9.09 to 11.12 MJ/kg DM, 61.30 to 75.54% and 90.35 to 138.05 g/kg DM, respectively. The ME, OMD and MP of common vetch was significantly (p<0.01) higher than those of the other hays due to low cell-wall contents and high crude protein. At the end of the experiment, differences in chemical composition of legume hays resulted in the differences in the in vitro gas production, gas production kinetics and the estimated parameters such as ME, OMD and MP. Common vetch can be recommended to hay producers and ruminant breeders, due to high ME, OMD and MP production.