Abstract: Tissue plasminogen activator (tPA) as a serine protease plays an important role in the fibrinolytic system and the dissolution of fibrin clots in human body. The production of this drug in plants such as tobacco could reduce its production costs. In this study, expression of tPA gene and protein targeting to different plant cell compartments, using various signal peptides has been investigated. For high level of expression, Kozak sequence was used after CaMV35S in the beginning of the gene. In order to design the final construction, Extensin, KDEL (amino acid sequence including Lys-Asp-Glu-Leu) and SP (γ-zein signal peptide coding sequence) were used as leader signals to conduct this protein into apoplast, endoplasmic reticulum and cytosol spaces, respectively. Cloned human tPA gene under the CaMV (Cauliflower mosaic virus) 35S promoter and NOS (Nopaline Synthase) terminator into pBI121 plasmid was transferred into tobacco explants by Agrobacterium tumefaciens strain LBA4404. The presence and copy number of genes in transgenic tobacco was proved by Southern blotting. Enzymatic activity of the rt-PA protein in transgenic plants compared to non-transgenic plants was confirmed by Zymography assay. The presence and amount of rt-PA recombinant protein in plants was estimated by ELISA analysis on crude protein extract of transgenic tobacco using a specific antibody. The yield of recombinant tPA in transgenic tobacco for SP, KDEL, Extensin signals were counted 0.50, 0.68, 0.69 microgram per milligram of total soluble proteins.
Abstract: Cadmium (II) (Cd) is one of the major toxic elemental
pollutants, which is hazardous for humans, animals and plants. γ-
Polyglutamic acid (γ-PGA) is an extracellular biopolymer produced
by several species of Bacillus which has been reported to be an
effective biosorbent for metal ions. The effect of γ-PGA on growth of
rice grown under laboratory conditions was investigated. Rice seeds
were germinated and then grown at 30±1°C on filter paper soaked
with Cd solution and γ-PGA for 7 days. The result showed that Cd
significantly inhibited the growth of roots, shoots by reducing root,
and shoot lengths. Fresh and dry weights also decreased compared
with control; however, the addition of 500 mg·L-1 γ-PGA alleviated
rice seedlings from the adverse effects of Cd. The analysis of
physiological traits revealed that Cd caused a decrease in the total
chlorophyll and soluble protein contents and amylase activities in all
treatments. The Cd content in seedling tissues increased for the Cd
250 μM treatment (P
Abstract: Whey is the lactose rich by-product of the dairy
industry, having good amount of nutrient reservoir. Most abundant
nutrients are lactose, soluble proteins, lipids and mineral salts.
Disposing of whey by most of milk plants which do not have proper
pre-treatment system is the major issue. As a result of which, there
can be significant loss of potential food and energy source. Thus,
whey has been explored as the substrate for the synthesis of different
value added products such as enzymes. β-galactosidase is one of the
important enzymes and has become the major focus of research due
to its ability to catalyze both hydrolytic as well as
transgalactosylation reaction simultaneously. The enzyme is widely
used in dairy industry as it catalyzes the transformation of lactose to
glucose and galactose, making it suitable for the lactose intolerant
people. The enzyme is intracellular in both bacteria and yeast,
whereas for molds, it has an extracellular location. The present work
was carried to utilize the whey for the production of β-galactosidase
enzyme using both yeast and fungal cultures. The yeast isolate
Kluyveromyces marxianus WIG2 and various fungal strains have
been used in the present study. Different disruption techniques have
also been investigated for the extraction of the enzyme produced
intracellularly from yeast cells. Among the different methods tested
for the disruption of yeast cells, SDS-chloroform showed the
maximum β-galactosidase activity. In case of the tested fungal
cultures, Aureobasidium pullulans NCIM 1050 was observed to be
the maximum extracellular enzyme producer.
Abstract: An attempt was made to study of nitrogen
components response of corn (Zea mays L.) to drought stress. A farm
research was done in RCBD as split-plot with four replications in
Khorramabad, west Iran. Drought stress levels as irrigation regimes
after 75 (control), 100, and 120 (stress) mm cumulative evaporation
were in main plots, and four seed corn varieties include 500 (medium
maturity), 647, 700, and 704 (long maturity) were as subplots.
Soluble protein, nitrate and proline amino acid were measured in
shoot and root at flowering stage, and grain yield was measured in
harvesting stage. As the drought progressed, the amount of nitrate
and proline followed an increasing trend, but soluble protein
decreased in shoot and root. The highest amount of nitrate and
proline was observed in longer maturity varieties than shorter ones,
but decrease yield of long maturity varieties was higher than medium
maturity varieties in drought condition, because of long duration of
stress.