*Author for Correspondence.

Abstract.
An experiment was carried out to investigate Single Nucleotide Polymorphisms (SNPs) in the coding regions of α-LA gene in two Indian buffalo breeds vis. Murrah (the best milk yielder) and South Kanara (used in agriculture operations). SNPs in α-LA potentially alter the gene expression and may be associated with differences in milk yield and quality. High molecular weight genomic DNA was extracted from 16 peripheral blood samples. Based on the 3090 bp sequences for bovine gene for alpha-lactalbumin (NCBI GenBank X06366) four primers were designed and used for amplification. The PCR products were electrophoresed, visualized and documented. Single Stranded Conformational Polymorphisms (SSCP) technique was used to screen for SNPs using the DCode Universal Mutation Detection System with 6% acrylamide gel. The individuals (samples) showing polymorphisms and also two samples per breed for each exon were further probed by DNA sequencing. DNA was eluted from the gel using a gel elution Kit. For sequencing, PCR reactions contained 5ul of eluted DNA and 10 pmoles of primer and 4ul of reaction mix. The sequencing products were analysed with the ABI Prism 377 DNA automated sequencer. In order to detect possible polymorphisms, the buffalo α-LA gene sequences observed were compared with the sequence for the same gene in NCBI gene bank AF194373. Sequence of exon 1 of all the animals were determined by direct sequencing. All the 8 buffaloes belonging to both the breeds showed a substitution at position 864 (C→T) and one South Kanara buffalo showed substitution at position 1264 (A→G). The polymorphism at position 864 (C→T) results in substitution of an amino acid P→S while the observed polymorphism at 1264 was a silent mutation in the coding region of the gene. No SNPs were observed in other three exons. The substitution observed in all the buffaloes concerned reveals Indian buffaloes which are riverine breeds differing from the buffaloes of China.

*Author for Correspondence.
Advanced Biotechnology and Breeding Centre, Malaysian Palm Oil Board (MPOB), P.O Box 10620, 50720 Kuala Lumpur.

Email: This email address is being protected from spambots. You need JavaScript enabled to view it.
Tel: 603: 8769 4501; Fax : 603-8926 1995

Abstract.
Restriction fragment length polymorphic (RFLP) probes derived from complementary DNA (cDNA) clones were developed for oil palm genome analysis. A total of 321 cDNA-RFLP probes were evaluated for their ability to detect polymorphism in a mapping family derived from the selfing of a Tenera guineensis palm (palm T128). Approximately 38% (123 probes) revealed polymorphism with at least one restriction enzyme. All the 123 informative probes were used to genotype the mapping family. Majority of the markers (80%) showed expected segregation ratios, indicating that most of the RFLP markers were inherited in a Mendelian manner. A total of 116 segregating markers were assigned to 20 linkage groups spanning 693cM. The RFLP markers were found to be largely well distributed and did not show excessive clustering in any particular region. This is the first published map for oil palm containing gene specific markers. The cDNA-RFLP probes mapped will not be merely anonymous markers with symbols, but point to the actual location of specific genes. The map also proved useful in revealing QTL associated with oil to wet mesocarp (O/WM) content.

*Author for Correspondence.
Department of Molecular Medicine
University of Malaya
50603, Kuala Lumpur, Malaysia

Phone: 603-79674718/7535
Fax: 603-79674957
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Abstract.
We had recently detected the germline D727E polymorphism of the thyroid-stimulating receptor (TSHR) gene in a cohort of patients with congenital hypothyroidism (CH). The role of TSHR genetic variants in CH has not been well studied. This study investigated the possible relationship between the polymorphism and CH in these patients. We studied 104 normal subjects as control and 33 patients with CH. A PCR-RFLP analysis was used to identify the mutation. There was no significant difference in either the genotype distribution or allelic frequencies between total CH and total controls for the D727E polymorphism (P=0.248 and 0.222 respectively). The controls and the patients with CH were further subdivided into 3 groups according to their ethnicities: Malay, Chinese and Indian. There were no significant differences in the D727E polymorphism genotype distribution between 1) the Malay patients with CH and the Malay normal healthy controls (P=0.442) 2) the Chinese patients with CH and the Chinese healthy controls (P=0.410) and 3) the Indian patients with CH and the Indian healthy controls (P=0.433). In addition, there were also no significant differences of the frequency of the C allele between 1) the Malay patients with CH and the Malay healthy controls (P=0.251) the Chinese patients with CH and the Chinese healthy controls (P=0.251). However, we found that the C allele for the D727E polymorphism was significantly more common in the Indian patients with CH as compared to the Indian healthy controls (P=0.03). These findings suggest that in this cohort of CH patients, the D727E polymorphism of the TSHR gene is a predisposing genetic factor for the development of CH in the Indian but not in the Malay and Chinese.

*Author for Correspondence.
School of Pharmacy and Health Sciences,
Faculty of Medicine, Pharmacy and Health Sciences,
International Medical University,
No. 126, Jalan 19/155B, Bukit Jalil,
57000 Kuala Lumpur, Malaysia
Tel: 60-3-86567228; Fax: 60-3-86567229;
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Abstract.
Dsn1p is a member of the MIND complex that forms part of the yeast kinetochore, which is essential for the proper chromosomal segregation during cell division. Its functionality is gene dosage dependent and it has characteristics of haploinsufficiency. Bioinformatics alignments predicted the existence of nuclear homologues in higher eukaryotic organisms. Literature on the possibility of Dsn1p being a functional homologue of these organisms is scarce. In this study we employed recombinant DNA expression technology to explore whether Dsn1p can function in a mammalian cell line, Chinese Hamster Ovary (CHO). Expression of rDsn1p in CHO cells induced cytopathic effects including changes in cellular morphology and cell size. Inhibition of cell growth was observed at the beginning the fourth post-transfection week. The recombinant CHO cell culture showed cytotoxic effects following the accumulation of the Dsn1p, resulting in apoptotic cell death; as evidenced by the presence of nuclear fragmentation and surface blebbing in the dying cells. This suggests that rDsn1p may interact with the counterpart/ligand of the nuclear homologue of this protein in CHO cells, resulting in nuclear anomalies and inhibition of cell growth, as observed in our previous study using yeast cells.