• 제목/요약/키워드: protein mixture

검색결과 764건 처리시간 0.031초

Construction of a High-Quality Yeast Two-Hybrid Library and Its Application in Identification of Interacting Proteins with Brn1 in Curvularia lunata

  • Gao, Jin-Xin;Jing, Jing;Yu, Chuan-Jin;Chen, Jie
    • The Plant Pathology Journal
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    • 제31권2호
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    • pp.108-114
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    • 2015
  • Curvularia lunata is an important maize foliar fungal pathogen that distributes widely in maize growing area in China, and several key pathogenic factors have been isolated. An yeast two-hybrid (Y2H) library is a very useful platform to further unravel novel pathogenic factors in C. lunata. To construct a high-quality full length-expression cDNA library from the C. lunata for application to pathogenesis-related protein-protein interaction screening, total RNA was extracted. The SMART (Switching Mechanism At 5' end of the RNA Transcript) technique was used for cDNA synthesis. Double-stranded cDNA was ligated into the pGADT7-Rec vector with Herring Testes Carrier DNA using homologous recombination method. The ligation mixture was transformed into competent yeast AH109 cells to construct the primary cDNA library. Eventually, a high qualitative library was successfully established according to an evaluation on quality. The transformation efficiency was about $6.39{\times}10^5$ transformants/$3{\mu}g$ pGADT7-Rec. The titer of the primary cDNA library was $2.5{\times}10^8cfu/mL$. The numbers for the cDNA library was $2.46{\times}10^5$. Randomly picked clones show that the recombination rate was 88.24%. Gel electrophoresis results indicated that the fragments ranged from 0.4 kb to 3.0 kb. Melanin synthesis protein Brn1 (1,3,8-hydroxynaphthalene reductase) was used as a "bait" to test the sufficiency of the Y2H library. As a result, a cDNA clone encoding VelB protein that was known to be involved in the regulation of diverse cellular processes, including control of secondary metabolism containing melanin and toxin production in many filamentous fungi was identified. Further study on the exact role of the VelB gene is underway.

역류분배를 이용한 모델단백질의 분리정제시 pH의 영향에 관한 연구 (Effects of pH on the Separation and Purification of Model Protein using Counter Current Distribution)

  • 이부용;이창호;이철호
    • 한국식품과학회지
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    • 제22권1호
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    • pp.56-60
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    • 1990
  • 역류분배 방법을 이용한 단백질의 분리정제 가능성을 높이기 위하여 4가지 모델단백질에 대하여 수용성 액체 2상계에서의 분획계수를 pH 변화에 따라 조사하였다. 또한 역류분배를 이용한 다중분배추출방식으로 모델 단백질의 혼합물을 수준의 pH(4.5-12.0)에서 분획하였다. 그 결과 pH에 따라 단백질들의 등전점 차이로 인한 분획계수(K)값이 다르게 나타서, 다중분배를 행할 때 각 단백질 분획들이 위치하는 튜브번호가 다르게 되므로 역류분배 방법을 통한 단백질의 분리 가능성을 보여주었다.

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Effects of microencapsulated organic acids on growth performance, nutrient digestibility, fecal microbial counts, and blood profiles in weaning pigs

  • Lee, Jun Soeng;Kim, Tae Heon;Song, Min Ho;Oh, Han Jin;Yun, Won;Lee, Ji Hwan;Kim, Yong Ju;Lee, Byong Kon;Kim, Hyeun Bum;Cho, Jin Ho
    • Journal of Animal Science and Technology
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    • 제63권1호
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    • pp.104-113
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    • 2021
  • This study was conducted to investigate the efficiency of a microencapsulated mixture of organic acids (MOA) with low protein in piglet feed on growth performance, diarrhea score, nutrient digestibility, fecal microbial counts, and blood profiles in weaning pigs. A total of 80 pigs [(Landrace × Yorkshire) × Duroc; 6.8 ± 0.48 kg] were randomly assigned to four dietary treatment groups: high protein (HP); low protein (LP); MOA1, LP + 0.2% MOA; and MOA2, LP + 0.3% MOA. The MOA2 group had higher average daily weight gains (during days 0-14 and days 0-28), diarrhea score (during days 0-14, during days 14-28 and days 0-28) and greater digestibility of dry matter (days 14 and 28) compared to the LP group (p < 0.05). However, there were no significant differences (p > 0.05) between the pigs fed diets with the MOA1 and MOA2 in blood profiles and fecal microflora. In conclusion, this study indicates that piglets fed 0.3% MOA in low protein diets maintained similar growth performance and nutrient digestibility, but alleviated the incidence of diarrhea compared to piglets fed high protein diets.

Functional Properties of Peptides in Mixed Whey and Soybean Extracts after Fermentation by Lactic Acid Bacteria

  • Dong-Gyu Yoo;Yu-Bin Jeon;Se-Hui Moon;Ha-Neul Kim;Ji-Won Lee;Cheol-Hyun Kim
    • Journal of Dairy Science and Biotechnology
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    • 제41권3호
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    • pp.113-125
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    • 2023
  • In this study, we explored the synergistic effects of whey protein concentrate (WPC) and soybean protein components after fermentation with lactic acid bacteria isolated from kimchi, and identified several peptides with desirable physiological functions, proteolysis, and immune effects. Antioxidant activity was determined using 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid, 1,1-diphenyl-2-picrylhydrazyl, ferric-reducing antioxidant power, and hydroxyl radical scavenging assays, followed by cross-validation of the four antioxidant activities. These assays revealed that samples with a 8:2 and 9:1 whey to soy ratio possessed higher antioxidant activity than the control samples. Antibacterial potency testing revealed high antibacterial activity in the 9:1 and 8:2 samples. Cytotoxicity testing of samples using 3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide revealed that only the 10:0, 1:9, and 0:10 samples had <80% viable cells, indicating no significant cytotoxicity. Nitric oxide (NO) assays revealed that NO expression was reduced in 8:2, 5:5, and 0:10 protein ratio fermentations, indicating low inflammatory reaction stimulatory potential. Cytokine expression was confirmed using an enzyme-linked immunosorbent assay kit. The 8:2 sample had the lowest inflammatory cytokine (interleukin [IL]-1α, IL-6, and tumor necrosis factor-α) levels compared with the lipopolysaccharide-treated group. Amino acid profiling of the 8:2 sample identified 17 amino acids. These results suggest that inoculating and fermenting Lactobacillus plantarum DK203 and Lactobacillus paracasei DK209 with an 8:2 mixture of WPC and soybean protein releases bioactive peptides with excellent anti-inflammatory and antioxidant properties, making them suitable for functional food development.

단백질(蛋白質) 최저요구량(最低要求量) 측정(測定)에 관한 연구(測定) (A Study on Estimation of Minimum Protein Requirement)

  • 왕수경;김미경
    • Journal of Nutrition and Health
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    • 제9권3호
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    • pp.8-17
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    • 1976
  • This study was carried out to estimate the minimum protein requirement for the Korean college girls, and to see the effect of carbohydrate intake on the minimum protein requirement in the rat. In the first experiment, three students attending Ewha Womans University were given protein free diet for 7 days while caloric intake were adjusted to maintain constant body weight. Endogenous nitrogen excretion was measured to estimate their minimum protein requirement. Second experiment was carried out in 3 parts to find out the effect of carbohydrate intake on the minimum protein requirement using 15 female, 50 days old rats weighing 120g. Inexperiment 2-1, the rats were fed protein free diet for 9 days and endogenous nitrogen excretion was measured to determine the minimum protein requirement of the rats. In experiment 2-2, the minimum casein requirement of the rats needed to maintain nitrogen equilibrium was estimated by the least squares regression method feeding 3 different levels of casein determined from the result of experiment 2-1. Finally, in experiment 2-3, the rats were given the minimum casein requirement for 7 days with 3 different levels of carbohydrate to find out the effect of carbohydrate intake on the nitrogen balance. The results of this study were as follows. Experiment I. Daily endogenous nitrogen losses of 3 college girls were 1.89g in urine, 1.10g in feces and 0.0108g from skin since the total daily endogenous nitrogen loss was 3.0g and the total daily nitrogen intake was 0.52g, the minimum nitrogen requirement of these subjects was about 2.48g/day (15.5g protein/day). Experiment II. Experiment 2-1: daily endogenous nitrogen losses of the rats fed protein free diet were $22.63{\pm}0.48\;mg$ through urine and $13.12{\pm}0.47\;mg$ in feces. The total daily endogenous nitrogen ouput was $35.71{\pm}1.27\;mg$ and the minimum protein requirement of the rats was 225mg/day in this study. Experiment 2-2: the minimum casein requirement needed to maintain the rats in nitrogen equilibrium was determined to be 340mg/day. Experiment 2-3: the rats were given 350mg/day casein with 3 different levels of carbohydrate (12,26 or 46 Cal by sugar-cornstarch mixture). The nitrogen balances were measured to be +0.0021, +0.0073 and +0.0143 respectively and there were significant differences among 3 groups.

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The effect of nanoemulsified methionine and cysteine on the in vitro expression of casein in bovine mammary epithelial cells

  • Kim, Tae-Il;Kim, Tae-Gyun;Lim, Dong-Hyun;Kim, Sang-Bum;Park, Seong-Min;Lim, Hyun-Joo;Kim, Hyun-Jong;Ki, Kwang-Seok;Kwon, Eung-Gi;Kim, Young-Jun;Mayakrishnan, Vijayakumar
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권2호
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    • pp.257-264
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    • 2019
  • Objective: Dairy cattle nutrient requirement systems acknowledge amino acid (AAs) requirements in aggregate as metabolizable protein (MP) and assume fixed efficiencies of MP used for milk protein. Regulation of mammary protein synthesis may be associated with AA input and milk protein output. The aim of this study was to evaluate the effect of nanoemulsified methionine and cysteine on the in-vitro expression of milk protein (casein) in bovine mammary epithelial cells (MAC-T cells). Methods: Methionine and cysteine were nonionized using Lipoid S 75 by high-speed homogenizer. The nanoemulsified AA particle size and polydispersity index were determined by dynamic light scattering correlation spectroscopy using a high-performance particle sizer instrument. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine the cytotoxicity effect of AAs with and without nanoionization at various concentrations (100 to $500{\mu}g/mL$) in mammary epithelial cells. MAC-T cells were subjected to 100% of free AA and nanoemulsified AA concentration in Dulbecco's modified Eagle medium/nutrient mixture F-12 (DMEM/F12) for the analysis of milk protein (casein) expression by the quantitative reverse transcription polymerase chain reaction method. Results: The AA-treated cells showed that cell viability tended to decrease (80%) in proportion to the concentration before nanogenesis, but cell viability increased as much as 90% after nanogenesis. The analysis of the expression of genetic markers related to milk protein indicated that; ${\alpha}_{s2}$-casein increased 2-fold, ${\kappa}$-casein increased 5-fold, and the amount of unchanged ${\beta}$-casein expression was nearly doubled in the nanoemulsified methionine-treated group when compared with the free-nanoemulsified methionine-supplemented group. On the contrary, the non-emulsified cysteine-administered group showed higher expression of genetic markers related to milk protein ${\alpha}_{s2}$-casein, ${\kappa}$-casein, and ${\beta}$-casein, but all the genetic markers related to milk protein decreased significantly after nanoemulsification. Conclusion: Detailed knowledge of factors, such nanogenesis of methionine, associated with increasing cysteine and decreasing production of genetic markers related to milk protein (casein) will help guide future recommendations to producers for maximizing milk yield with a high level of milk protein casein.

중부지역 초지에서 혼파조합 및 유기질 비료 시용이 목초 수량, 식생 구성 및 사료가치에 미치는 영향 (Effect of Seed Mixture and Organic Fertilizer Application on Productivity, Botanical Composition and Forage Quality in Middle Mountainous Pasture)

  • 김종근;이우위;김맹중;김학진;최승기;김종덕
    • 한국초지조사료학회지
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    • 제37권4호
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    • pp.337-344
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    • 2017
  • 본 시험은 우리나라 중부지역(충남 금산, 해발 250m)에서 유기물 시용과 및 혼파조합에 따른 신규조성 초지의 생산성 변화를 평가하기 위하여 수행하였다. 총 6개의 처리 (오차드 그라스 위주, 티머시 위주 그리고 톨페스큐 위주의 혼파 초지에 유기질 비료의 시용 유무)를 두고 2014년 가을에 조성하여 2015년부터 2016년까지 조사하였다. 초장은 오차드그라스 위주 혼파조합에서 길었으며 2차 수확시는 오차드그라스 위주 및 톨페스큐 위주의 혼파조합에서 길었다. 건물함량은 대체적으로 처리간에 유의적인 차이가 없었다(P<0.05). 식생은 1차 수확시 오차드그라스 위주 혼파조합에서 잡초의 비율이 높았으며 3차 수확시에는 모든 처리구에서 두과목초가 사라졌다. 생초 및 건물수량은 1년차보다는 2년차에서 더 높게 나타났다. 또한 1년차에 유기물을 시용한 오차드그라스 위주 혼파조합에서 유의적으로 높았으며(P<0.05), 2년차에서는 유기물을 시용한 톨페스큐 위주 혼파조합에서 가장 높게 나타났다. 조단백질 함량은 톨페스큐 혼파조합에서 높게 나타났으며 티머시 위주 혼파조합에서 ADF 및 NDF 함량이 낮았고 소화율은 높았다. 조성 2년차에서는 2회 및 4회 예취시 목초의 사료가치가 높게 나타났으며 평균 RFV값도 4차에서 가장 높았다. 이상의 결과를 종합하여 볼 때 중부지역 구릉지 초지 조성시 생산성 면에서는 유기물을 시용한 오차드그라스와 톨페스큐 위주 혼파조합이 우수하였으며 사료가치적 측면에서는 티머시 위주 혼파조합이 우수한 것으로 나타났다.

혈청단백질분획(血淸蛋白質分劃)에 대한 연구(硏究) (I, II) (Studies on Bovine Serum Protein Fractions (I, II))

  • 임봉호
    • 대한수의학회지
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    • 제5권1호
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    • pp.1-16
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    • 1965
  • I. A Comparison of Sodium Sulfate Precipitation and Zone(Paper, Agar) Electrophoresis; Many kinds of techniques have been used for fractionating serum proteins. In the present study, using bovine serum, the fractions obtained with sodium sulfate were compared with those determined by zone electrophoresis. 1. Fibrinogen was precipitated with 4 to 10 percent of sodium sulfate. 2. ${\gamma}$-globulin required 10 to 16 percent of the salt for precipitation. 3. ${\beta}$-globulin began to precipitate at 12 percent sodium sulfate, and completed precipitation at approximately 26 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. 4. ${\alpha}$-globulin completed precipitation at 13 to 28 percent sodium sulfate in paper electrophoresis and at 22 percent in agar electrophoresis. 5. Albumin began to precipitate at 14 percent of the salt, and was free from the mixture of globulins approximately at 28 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. The results of comparing fractions by the two methods were as follows: 1. Euglobulin (15%) was equal to the sum of the most ${\gamma}$-globulin and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. 2. Pseudoglobulin I (15-17.5%) corresponded to the most ${\alpha}$-, ${\beta}$-globulins and a small quantity of albumin. 3. Pseudoglobulin II(18-22%) was a mixture of the ${\alpha}$-, ${\beta}$-globulins and albumin fraction. 4. Albumin (above 22%) contained the most albumin fraction separated by zone electrophoresis and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. As mentioned above the fractions obtained with sodium sulfate were a mixture of the various proportion of the fractions determined by zone electrophoresis. The solubility of serum fractions to sodium sulfate coincided with the mobility of those by zone electrophoresis. (By percent of sodium sulfate we mean gram of sodium sulfate contained in $100m{\ell}$ of solution). II. Immunological Studies on Serum Protein Fractions with Sodium Sulfate; In the previous report the fractions of bovine serum protein with sodium sulfate compared with those obtained by zone electrophoresis, and the findings were that the former contained various proportion components of the latter. In this study the author studied whether or not the fractions with sodium sulfate are simple component antigenically by immunoelectrophoresis and micro double diffusion test (Immuno-precipitation), using rabbit antiserum to bovine serum. In immunoelectrophoresis, normal bovine serum developed with rabbit antibovine serum showed about ten distinct precipitin arcs. The distribution of these arcs was as follows: 1 albumin, 2 ${\alpha}_1$-, 3 ${\alpha}_2$-, 2 ${\beta}_1$-, ${\beta}_2$-, and 1 ${\gamma}$-globulin (Fig. 7, 9). In micro double diffusion test, five to six precipitation bands could be seen between antigens and antibody, the order of the precipitation bands location is albumin, ${\alpha}$-, ${\beta}$-, and ${\gamma}$-globulin from the side of antiserum well (Fig.19). Frequently the ${\alpha}$-, and ${\beta}$-precipitation bands were separated into two or three precipitation bands, which indicated that these globuline are not a pure component antigenically as shown in immuno-electrophoresis. In both Immunological methods, the two ${\alpha}$-, ${\beta}$-precipitin arcs and bands appeared clear and strong, indicating that the two globulins reacted as strong antigens. The precipitate reaction of ${\gamma}$-globulin was shown at 12 to 16 percent sodium sulfate; ${\beta}$-globulin at 12 to 20 percent; ${\alpha}$-globulin at 12 to 22 percent (immuno-electrophoresis), at 12 to 26 percent (Diffusion); and albumin at above 22 percent. Antigenically euglobulin contained ${\gamma}$-, ${\beta}$-, and ${\alpha}$-globulins, Pseudoglobulin I and Pseudoglobulin II were composed of ${\alpha}$-, and ${\beta}$-globulins, and albumin was a mixture of ${\alpha}$-globulin and albumin determined by zone electrophoresis. The results indicated that the fractions of serum protein obtained by either method were constituents of various proteins antigenically except ${\gamma}$-globulin and albumin by Zone electrophoresis.

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친환경 소재 잣나무 목재와 케나프 줄기 혼합물의 항산화 및 미백효과 (Antioxidant Activities and Whitening Effects of a Mixture of the Eco-friendly Materials Pinus koraiensis and Hibiscus cannabinus L.)

  • 오민정;염현지;채정우;이진영
    • 생명과학회지
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    • 제31권3호
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    • pp.305-313
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    • 2021
  • 본 연구는 친환경 소재인 잣나무 목재 추출물과 케나프 줄기 추출물의 항산화 및 미백 효능을 검증하고, 잣나무 목재 추출물과 케나프 줄기 추출물을 동일 비율로 혼합하였을 때 유효성분들의 상호작용을 확인하고자 하였다. 각각의 추출물을 1:1의 비율로 혼합한 추출물의 전자공여능 및 ABTS 소거능을 측정한 결과 1,000 ㎍/ml에서 추출물 성분들의 상승작용을 통해 혼합 추출물이 93.7%, 94%로 가장 높은 효능을 나타내었다. 잣나무 목재, 케나프 줄기 및 혼합 추출물의 tyrosinase 억제활성을 측정한 결과 1,000 ㎍/ml 농도에서 40%, 27.5%, 43%의 효과를 보였다. 따라서 추출물에 함유되어 있는 유효 성분들의 상승효과로 혼합 추출물의 높은 효능을 확인할 수 있었다. 잣나무 목재, 케나프 줄기 및 혼합 추출물의 melanoma cell에 대한 세포 독성 측정 결과 1,000 ㎍/ml 농도에서 각각 87.4%, 80.2%, 98%의 생존율을 나타내었으며, 결과적으로 혼합 추출물은 길항작용을 통해 같은 농도에서도 100%에 가까운 세포 생존율을 보였다. 잣나무 목재와 케나프 줄기 혼합 추출물의 Western blot에 의한 MITF, TRP-1, TRP-2 및 tyrosinase의 단백질 발현 효과를 측정한 결과, 100 ㎍/ml 농도에서 53.9%, 64.8%, 67.3%, 56.1%로 단백질 발현 억제 효과를 보였다. 또한 RT-PCR을 통해 혼합 추출물이 미백관련 인자들의 mRNA 발현에 미치는 억제효과를 측정한 결과 100 ㎍/ml 농도에서 각각 54.4%, 64.9%, 66.6%, 63.1%로 발현이 감소함을 확인할 수 있었다. 그 결과 잣나무 목재와 케나프 줄기를 1:1 비율로 혼합한 추출물의 항산화 및 미백 효능을 검증하였으며, 본 연구결과를 종합하였을 때 잣나무 목재와 케나프 줄기를 혼합한 추출물이 기능성 화장품 소재로서의 활용 가능성을 확인할 수 있었다.

Nutritive Value and Utilization of Perennial Grasses Intercropped with Soybean Fodder by Crossbred Heifers in Humid-subtropics of Himachal Pradesh

  • Radotra, Sudesh;Katoch, B.S.
    • Asian-Australasian Journal of Animal Sciences
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    • 제15권12호
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    • pp.1754-1759
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    • 2002
  • A study was carried out to investigate the nutritive value and utilization of hybrid sorghum and perennial grass species viz. setaria (Setaria anceps) and hybrid napier when intercropped with soybean by growing Jersey crossbred heifers. Fifteen growing crossbred heifers (Jersey${\times}$Red Sindhi) of between 7-10 months age and pre-trial average body weight of 49-50 kg were divided on the basis of weight in to three treatment groups viz. $T_1$-hybrid sorghum+soybean, $T_2$-setaria+soybean and $T_3$-hybrid napier+soybean in a completely randomized block design. Intercropped forages were harvested fresh, chaffed and mixed before they were offered to the heifers. Chemical composition of the herbage, dry matter intake (DMI), body weight gain and nutrient digestibility co-efficients were estimated. The herbage mixtures had crude protein (CP) content in the range of 11.87 to 13.86% and ether extract (EE) contents were 2.91 to 3.11%, respectively. The herbage mixtures were rich in minerals (ash). The gross energy (kcal/g DM) was higher in hybrid napier+soybean, while hybrid sorghum+soybean and setaria+soybean herbage mixtures had lower value for gross energy. The hybrid sorghum+soybean and setaria+soybean herbage mixtures had higher contents of NDF, ADF, cellulose, lignin and silica as compared to that of hybrid napier+soybean herbage mixture. The heifers fed hybrid napier+soybean herbage mixture had significantly (p<0.05) higher $DMI\;g/kg\;W^{0.75}$ ($97.41{\pm}4.34$) as compared to hybrid sorghum+soybean ($88.31{\pm}2.66$) and setaria+soybean ($79.29{\pm}1.06$) herbage mixtures. Nutrients digestibility, DCP percent, DCP intake and nitrogen balance were significantly (p<0.05) higher in the heifers fed on hybrid napier+soybean herbage mixture. There was a significant (p<0.05) difference among different herbage mixtures in TDN. The heifers on setaria+soybean herbage mixture had lower average body weight gain (g/day) than those on hybrid sorghum+soybean and hybrid napier+soybean herbage mixtures. Data obtained in this experiment demonstrated that herbage mixture of hybrid napier+soybean was better than hybrid sorghum+soybean and setaria+soybean herbage mixtures in the nutrition of growing heifers. It had highest nutritive value, better digestibility co-efficients which showed better growth rate and higher feed efficiency. In ranking, hybrid napier+soybean herbage mixture was better followed by hybrid sorghum+soybean and setaria+soybean in nutritive value in the parameters studied. For future wasteland development program in humid-sub tropics of Himachal Pradesh hybrid napier and its intercropping with soybean is recommended for general adoption because of its better adaptability and higher nutritive value.