Biotecnologia Aplicada, vol. 16, no. 02, 1999
BIOSCI Administrator
CC Vol. 16 No. 2 (April-June, 1999)
CC ISSN 0864-4551 (printed)
CC ISSN 1027-2852 (electronic)
CC Copyright 1999, Elfos Scientiae.
CC Founded in 1983 and published as Interferon y
CC Biotecnologia until 1990, Biotecnologia Aplicada is a
CC peer-reviewed scientific journal. It appears four times
CC per year, sponsored by the Ibero-Latin-American Society
CC of Biotechnology Applied to Health.
CC BIOTECNOLOGIA APLICADA is a vehicle of Iberian-Latin-
CC American scientists working in different fields of modern
CC biotechnology for the diffusion of their experiences and
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CC a) Molecular genetics and recombinant DNA technology;
CC b) Therapeutic and prophylactic uses of molecules
CC obtained by biotechnological methods;
CC c) Immunotechnology and development of diagnostic
CC methods;
CC d) Chemistry, biochemistry and design of molecules with
CC biological activity;
CC e) Application of recombinant DNA techniques and its
CC products in agriculture and animal science;
CC f) Transgenesis;
CC g) Fermentation, recovery and purification of biotech-
CC nological products;
CC h) Scaling-up, plant design and economic evaluation;
CC i) Control and automation of processes.
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CC Ave. 31 entre 158 y 190,
CC Cubanacan, Playa
CC Ciudad de La Habana,
CC Apdo. 6072, Habana 6, Cuba.
CC Telephones: (53-7) 33 1917 / 21 8466 / 21 8164
CC Fax: (53-7) 33 1917 / 21 8070 / 33 6008
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AU Javier Menéndez
IN División de Biotecnología Industrial. Centro de Ingeniería
Genética y Biotecnología. AP 6162, CP 10600. Ciudad de La
Habana, Cuba. Fax: (53-7) 33 6008;
E-mail: yeas...@cigb.edu.cu
TI Piruvato-carboxilasa de levadura
DE pyruvate carboxilase
DE promoters
DE PYC1
DE PYC2
DE yeast
SO Biotecnología Aplicada 1999;16:75-82
AB Pyruvate carboxylase plays an important role in
intermediary metabolism, catalyzing the formation of
oxaloacetate from pyruvate and HCO3-. It thus provides
oxaloacetate for gluconeogenesis and for replenishing
tricarboxylic acid cycle for fatty acid, amino acid and
neurotransmitter synthesis. The enzyme is highly conserved
and it is found in a great variety of organisms including
bacteria, yeasts, fungi and plants, as well as in higher
organisms. The yeast Saccharomyces cerevisiae contains two
structural genes for pyruvate carboxylase, which are
regulated in different ways. On the other hand, Pichia
pastoris has only one gene. This protein is a member of a
group of biotin-dependent enzymes and the disruption of
their coding genes results in the incapacity of the yeast
to grow on a minimal medium containing glucose as the sole
carbon source. This phenotype led to the isolation of
mutants from these two yeasts, which are affected in the
glucose repression of a series of genes
CC Review article
CC Language: Spanish
AU Raúl Gómez, Joel Madrazo, Javier González, Glay Chinea,
Alexis Mussachio, Armando Rodríguez, Gabriel Padrón
IN División de Química-Física, División de Vacunas. Centro de
Ingeniería Genética y Biotecnología. AP 6162, Ciudad de La
Habana, Cuba. Telf: (53-7) 21 8164; Fax: (53-7) 33 6008;
E-mail: raul....@cigb.edu.cu
TI Caracterización estructural y funcional de la proteína
recombinante P64k de Neisseria meningitidis
DE cross-linking
DE dihydrolipoamide-dehydrogenase
DE flavoprotein
DE Neisseria meningitidis
SO Biotecnología Aplicada 1999;16:83-87
AB The recombinant protein P64k from the bacterium Neisseria
meningitidis has been identified as a dihydrolipoamide
dehydrogenase. The enzyme catalyses the stoichiometric
oxidation of dihydrolipoamide by nicotinamide adenine
dinucleotide, involving a flavin adenine dinucleotide as
cofactor. Chemical modification with iodoacetamide
indicated the involvement in catalysis of a reversibly
reducible disulphide bond. Migration of cross-linked
protein in polyacrylamide gel electrophoresis showed that
the protein is a functional homodimer. Gel filtration
chromatography showed the P64k protein to migrate beyond
the expected molecular weight. This abnormal behaviour is
discussed taking into account the structure-function
relationship
CC Research article
CC Language: Spanish
AU Juan Roca, Masood Ahmad, Subrat Kumar Panda, Guillermo
Padrón, Shahid Jameel
IN Division of Diagnostics and Immunotechnology. Center for
Genetic Engineering and Biotechnology. PO Box 6162,
Havana 10600, Cuba. Fax: (53-7) 21 8008;
E-mail: di...@cigb.edu.cu. Virology Group. International
Center for Genetic Engineering and Biotechnology.
PO Box 10504, Aruna Asaf Ali Marg, New Delhi 110067,
India. Department of Pathology. All India Institute of
Medical Sciences, New Delhi, India
TI Hepatitis C Virus Genotyping in Developing Countries:
Results from Cuba, India and Turkey
DE HCV genotypes
DE hepatitis C virus
DE PCR
SO Biotecnología Aplicada 1999;16:88-92
AB Infection by hepatitis C virus (HCV) results in liver
disease with a high rate of associated chronicity.
Significant genetic variation is seen among HCV isolates
based on nucleotide sequence homologies, which has allowed
grouping into a number of genotypes. However, most of the
information about HCV genotypes is based on studies from
developed countries with less information available from
developing countries. Here we present the results of HCV
genotype determinations in 128 sera from 74 patients in
three countries: Cuba, India and Turkey. An established
PCR-based genotyping method was optimized to accurately
detect multiple genotypes in a given sample. Type II (1b)
HCV, which correlates with more aggressive forms of the
disease and lower response rates to interferon was most
commonly found in the patients from the countries studied.
While the majority of patients (58.1%) were infected with
a single genotype, dual infections were also found
frequently (28.4%). This report discusses the utility of a
genotyping assay and the prevalence of genotypes
CC Research Article
CC Language: English
AU Juan P Martínez-Soriano, Norma E Leyva-López, María E
Zavala-Soto, Marie Bères, Diana S Leal-Klevezas
IN Centro de Investigación y de Estudios Avanzados del
Instituto Politécnico Nacional. Unidad de Biotecnología e
Ingeniería Genética de Plantas. AP 629, Irapuato,
Gto. México. E-mail: jp...@irapuato.ira.cinvestav.mx
Institut National Agronomique Paris-Grignon. 16, rue
Claude Bernard 75231 Paris Cedex 05, Francia. Centro de
Investigación Biomédica del Noreste, Instituto Mexicano
del Seguro Social. San Luis Potosí y 2 de abril, Colonia
Independencia, Monterrey, NL México
TI Detección molecular del agente causal del síndrome
"bola de hilo" de la papa en semillas infectadas y
asintomáticas
DE diagnosis
DE mycoplasma-like organisms
DE PCR
DE phytoplasmas
SO Biotecnología Aplicada 1999;16:93-96
AB Economic losses due to the potato diseases caused by
phytoplasmas are of considerable value in Mexico (up to
100% in some cases). "Punta morada" and "bola de hilo"
(local names meaning "purple top" and "ball of twine",
respectively) are the main syndromes observed in the
field. The latter is one of particular interest among
potato seed producers because it affects tuber germination
when infected tubers are used as seed. Because
phytoplasmas cannot be cultured in vitro and their
detection by visual inspection, by serological methods or
by electron microscopy is inefficient and performed late
during disease development, disease eradication is an
impossible task. In this report, PCR was used as a new
tool to detect early or latent phytoplasma infections in
symptomless potato "seeds", which will eventually produce
"hair sprouts"
CC Research Article
CC Language: Spanish
AU Carmen E Gómez, Javier J Menéndez, Bianca M García
IN División de Biotecnología Industrial, Centro de Ingeniería
Genética y Biotecnología. AP 6162, CP 10600, Ciudad de La
Habana, Cuba. Teléfonos: (53-7) 21 6022, 21 8164,
Fax: (53-7) 21 8070. E-mail: dextr...@cigb.edu.cu
TI Expresión del gen dex en la levadura Kluyveromyces lactis
DE dextranase
DE gene expression
DE Kluyveromyces lactis
DE Penicillium minioluteum
SO Biotecnología Aplicada 1999;16:97-102
AB In this study, the expression and secretion of dextranase
enzyme from the fungus Penicillium minioluteum in the
non-conventional yeast Kluyveromyces lactis, was
evaluated. The cDNA encoding this protein with its own
signal peptide was integrated into the genome of K. lactis
strain MW105-2A under the control of the LAC4 promoter.
Dextranase was detected by the hydrolysis of blue dextran
added to solid growth medium and the enzymatic activity
was determined colorimetrically by the DNSA method.
Dextranase was efficiently expressed and secreted in this
yeast. The obtention of this construction constitutes the
first step in the use of dextranase-encoding gene as
reporter in gene expression studies in the yeast K. lactis
CC Research Article
CC Language: Spanish
AU Antonieta M Herrera, Dania Vázquez, Leonor Navea,
Leonor Lobaina, Diógenes Quintana, Annara Nápoles,
Yenela García, Carlos A Duarte
IN Vaccine Division, AIDS Department. Center for Genetic
Engineering and Biotechnology. PO Box 6162, Havana 10600,
Cuba. Phone: (53-7) 21 8008, 21 8466.
Fax: (53-7) 21 8070, 33 6008 E-mail: amhe...@cigb.edu.cu
Laboratorio de Investigaciones de SIDA, San José,
La Habana, Cuba
TI Effect of Different Adjuvants and Immunomodulators on the
Humoral Immune Response of Rabbits and Mice against
HIV-1-derived Multi-epitope Polypeptides
DE adjuvants
DE HIV
DE immunomodulators
DE vaccines
DE V3 loop
SO Biotecnología Aplicada 1999;16:103-108
AB The third variable region (V3 loop) of the human
immunodeficiency virus (HIV) external glycoprotein gp120
contains the principal neutralizing domain of this
protein. Our group has developed multi-epitope
polypeptides (MEP), bearing several copies of the V3 loop
from different HIV-1 isolates. These chimeric proteins
have been able to elicit broadly reactive neutralizing
antibodies when administered in Complete Freund's Adjuvant
(CFA). For human vaccines, a less reactogenic adjuvant is
required. The MEPs TAB9 and TAB13 contain the V3 region
from six and eight HIV-1 isolates, respectively, fused to
the amino terminus of the Neisseria meningitidis P64K
protein. In this paper we describe the effect of several
adjuvants and immunomodulators on the antibody response
against these MEPs in rabbits and mice. Oil adjuvants
proved to be more efficient in promoting the antibody
response against MEPs than Alum, Quil A or combinations of
Alum with IL-2 and gIFN. The subclass composition of the
antibody response was very dependent on the adjuvant
employed. CFA induced high levels of IgG2a and IgG2b,
while for the rest of the products IgG1 was predominant.
We also concluded that the novel oil adjuvant Montanide
ISA720 is as efficient as CFA or Incomplete Freund's
Adjuvant in stimulating the humoral response in mice and
rabbits and therefore, it was selected for further studies
in primates.
CC Research Article
CC Language: English
AU Thelvia I Ramos Gómez, Estela Morales Peralta, Teresa
Collazo Mesa, Suany Ojeda Fernández, Aida Bertoli Avella,
Luis Heredero Baute
IN Centro Nacional de Genética Médica. Centro colaborador de
la OMS para el desarrollo de enfoques genéticos en la
promoción de la salud. Instituto Superior de Ciencias
Médicas de La Habana, Cuba. Ave 31 No. 3102, Playa 16,
Ciudad de La Habana, Cuba. Teléfono: (53-7) 21 9511,
extensiones 268 y 361 Fax: (53-7) 24 6257;
E-mail: cng...@genmed.giron.sld.cu
TI Presencia de la mutación G1138A del gen del R3FCF en un
grupo de pacientes acondroplásicos cubanos
DE achondroplasia
DE autosomal dominant
DE FGFR3
DE hipoachondroplasia
DE osteochondrodysplasias
DE thanatophoric dysplasia
SO Biotecnología Aplicada 1999;16:109-111
AB Achondroplasia, inherited as an autosomal dominant trait,
is the most common human skeletal dysplasia. Prevalence at
birth in Cuba was estimated in 1/25 000. Achondroplasia
locus was assigned to chromosome 4p16.3 and includes the
coding region for fibroblast growth factor receptor 3
(FGFR3). Two point mutations in exon 10 of the FGFR3 gene
(G1138A and G1138C) have been described. These mutations
create new restriction sites for Sfc1 (the most frequent
mutation) and Msp1. Genomic DNA isolated from blood
samples of 40 Cuban patients (24 relatives and 16 sporadic
cases) was studied. A region of 164 bp that includes the
transmembrane domain of FGFR3 was amplified by PCR and
the amplicons were analized with the restriction enzimes
Sfc1 and Msp1. All affected individuals showed fragment
sizes of 109 and 55 bp corresponding to the digestion by
Sfc1; therefore, they had the same mutation G1138A
CC Research Article
CC Language: Spanish
AU Mayte Nerey Olivares, Rolando Ochoa Azze, Juan C Martínez
Rodríguez, Tania Licea Verdecia, Xenia Ferriol Marchena,
Ana M García Malberti, Rosa Blanco González,
Eric Estrada González
IN DACTA, Instituto "Finlay". Ave 27 No 19805. AP 16017,
CP 11600, La Lisa, Ciudad de La Habana, Cuba.
Fax: (53-7) 33 6754; E-mail: oc...@finlay.edu.cu
TI Validación de un ELISA para la cuantificación de IgG
humana anti-polisacarido capsular de Neisseria
meningitidis serogrupo C
DE anti-polysaccharide C ELISA
DE Neisseria meningitidis
DE polysaccharide C
SO Biotecnología Aplicada 1999;16:113-115
AB An indirect ELISA against capsular polysaccharide of
Neisseria meningitidis serogroup C was developed to
evaluate the immune response to this component of
VA-MENGOC-BC(r) Cuban vaccine. Polystyrene plates coated
with poly-L-lysine and polysaccharide antigen ("Finlay"
Institute), are first incubated with human serum samples.
Antibodies are detected by addition of anti-human
IgG/alkaline phosphatase conjugate, which recognizes
antibodies against polysaccharide C. The enzymatic
reaction is evidenced by processing of the specific
substrate p-nitrophenylphosfate. The detection limit of
the assay is 367 U/mL of specific IgG in human serum.
Intra- and interassay lack of precision was below 10%.
Parallelism, linearity and recovery were ± 10% of the
expected values. The relation between the standard of the
"Finlay" Institute and the PB2 standard from the Center
for Disease Control (CDC), Atlanta, USA, was determined.
The regression equation was CDC = 0.5398 Finlay +143
CC Article on Techniques
CC Language: Spanish
AU Javier Menéndez
IN División de Biotecnología Industrial, Centro de Ingeniería
Genética y Biotecnología. AP 6162, CP 10600. Ciudad de La
Habana, Cuba. Fax: (53-7) 21 8070.
E-mail: yeas...@cigb.edu.cu
TI Conventional and Non-conventional Yeasts in Modern
Biotechnology
SO Biotecnología Aplicada 1999;16:117-121
CC Report
CC Language: English
AU José A Cremata
IN División de Biotecnología Industrial, Centro de Ingeniería
Genética y Biotecnología. AP 6162, CP 10600. Ciudad de La
Habana, Cuba. Fax: (53-7) 21 8070.
E-mail: bio...@cigb.edu.cu
TI Structural Analysis of Glycoproteins. Its Importance
SO Biotecnología Aplicada 1999;16:121-125
CC Report
CC Language: English
AU Peter Aleström, José de la Fuente
IN Department of Biochemistry, Physiology and Nutrition,
Norwegian College of Veterinary Medicine. PO Box 8146 DEP,
N-0033 Oslo, Norway. División de Genética de Células de
Mamíferos, Centro de Ingeniería Genética y Biotecnología.
AP 6162, CP 10600, Ciudad de La Habana, Cuba.
Telef: (53-7) 21 8164; Fax: (53-7) 21 8070;
E-mail: jose.de...@cigb.edu.cu
TI Genetically Modified Fish in Aquaculture: Technical,
Environmental and Management Considerations
DE aquaculture
DE fish
DE safety
DE transgenic
SO Biotecnología Aplicada 1999;16:127-130
AB Genetically modified (GM) fish offer new possibilities for
the improvement of production in aquaculture. It allows
the introduction of novel traits or the improvement of old
ones, in such a way that is out of reach for classical
selection breeding. Examples of genes with commercial
potential are among those which control growth, disease
resistance, freeze tolerance, sexual maturation, food
quality and food preservation parameters. Consumption of
GM fish does not represent a health risk in principle. The
safety of GM food is dependent on the character of the
transgene, the transgene product and the new phenotype.
Ethics and animal protection concerns demand the
development of healthy fish only. Environmental safety
calls for efficient biological containment in order to
minimize possible effects caused by released farm animals.
Improvements of disease control will support both
production economy and the environment, in case of
escapes. Since aquaculture includes both marine and fresh
water species, it can be developed as new food production
strategies in most countries all over the world. To avoid
large-scale technology transfer failures, it is important
to adapt to the regional and local needs. This calls for
international research collaboration aiming at regional
and local competence development sufficient for the
technology implementation
CC Focus
CC Language: English