Detection of Polymorphism of Growth Hormone Gene for the Analysis of Relationship between Allele Type and Growth Traits in Karan Fries Cattle

Pal, Aruna,Chakravarty, A.K.,Bhattacharya, T.K.,Joshi, B.K.,Sharma, Arjava Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.10

The present study was conducted to detect polymorphism at growth hormone gene in Karan Fries bulls. A 428 bp fragment of growth hormone gene spanning over $4^{th}$exon, $4^{th}$intron and $5^{th}$ exon was amplified and digested with AluI restriction enzyme to identify polymorphism at this locus. Karan Fries bulls were found to be polymorphic at this locus. Two genotypes LL and LV were identified in Karan Fries with higher allelic frequency for L allele. In Karan Fries males, the average birth weight, 3 months body weight and daily body weight gains of LL homozygotes were significantly higher than that of LV heterozygotes. Genetic distances of KF bulls with respect to genotype along with 3 months body weight and average daily body weight gain forms a single cluster of bulls with LL genotype, while individuals with LV genotype forms three distinct clusters indicating more influence of L allele on growth traits.

Association of Beta-lactoglobulin Polymorphism with Milk Production Traits in Cattle

Badola, S.,Bhattacharya, T.K.,Biswas, T.K.,Kumar, Pushpendra,Sharma, Arjava Asian Australasian Association of Animal Productio 2003 Animal Bioscience Vol.16 No.11

The study was carried out in Sahiwal, Holstein Friesian, Jersey and crossbred cattle to find out the effect of genotype of beta-lactoglobulin gene on milk production traits. The polymorphism at beta-lactoglobulin gene was identified by conducting PCRRFLP studies. A 398 bp fragment of the gene was amplified and digested with Hae III restriction enzyme. The two alleles A and B and three genotypes AA, AB and BB were identified in all cattle breeds. The frequency of B allele was comparatively higher than that of A allele. The AA genotype produced significantly higher milk yield in Sahiwal cattle whereas BB genotype yielded higher milk in Holstein friesian cattle. In other cattle breeds the genotypic effect was non-significant. In conclusion it may be stated that the genotype with significantly higher milk yield may be favoured in the farm along with other conventional selection criteria to enhance the milk production of animals.

DNA Polymorphism in SLC11A1 Gene and its Association with Brucellosis Resistance in Indian Zebu (Bos indicus) and Crossbred (Bos indicus×Bos taurus) Cattle

Kumar, Nishant,Ganguly, Indrajit,Singh, Rajendra,Deb, Sitangsu M.,Kumar, Subodh,Sharma, Arjava,Mitra, Abhijit Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.7

The PCR- restriction fragment length polymorphism (RFLP) in and around TM4 of SLC11A1 gene and its association with the incidences of brucellosis in Hariana breed (Bos indicus) and Holstein Friesian crossbred (Bos indicus${\times}$Bos taurus) cattle was examined. A fragment of 954 bp encoding the TM4 was amplified, and RFLP was identified by digestion of the amplicon independently with AluI and TaqI. The amplicon (GenBank Acc. No. AY338470 and AY338471) comprised of a part of exon V (<59 bp) and VII (62>), and entire intron 5 (423 bp), exon VI (71 bp) and intron 6 (339 bp). Digestion with AluI revealed the presence of two alleles viz, A (281, 255, 79 and 51 bp) and B (541, 255, 79 and 51 bp). The frequency of A allele was estimated as 0.80 and 0.73 in Hariana and crossbred cattle, respectively. Due to presence of a polymorphic TaqI site at intron 5, two alleles: T (552 and 402 bp) and Q (231, 321 and 402 bp) were identified. The frequency of T allele was estimated as 0.96 and 0.97, respectively. For association study, on the basis of serological tests and history of abortion, the animals were grouped into "affected" and "non-affected". However, no association could be established with the observed RFLPs.

Characterization of MHC DRB3.2 Alleles of Crossbred Cattle by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism

Paswan, Chandan,Bhushan, Bharat,Patra, B.N.,Kumar, Pushpendra,Sharma, Arjava,Dandapat, S.,Tomar, A.K.S.,Dutt, Triveni Asian Australasian Association of Animal Productio 2005 Animal Bioscience Vol.18 No.9

The present investigation was undertaken to study the genetic polymorphism of the DRB3 exon 2 in 75 crossbred cattle by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Five genotypes i.e. HaeIII-a, HaeIII-b, HaeIII-e, HaeIII-ab and HaeIII-ae were observed when the 284 bp PCR products were digested with HaeIII restriction enzyme. The corresponding frequencies of these patterns were 0.53, 0.04, 0.01, 0.38 and 0.04, respectively. Digestion with RsaI restriction enzyme resolved 24 different restriction patterns. The frequencies of these patterns ranged from 0.013 (RsaI-f, RsaI-k and RsaI-c/n) to 0.120 (RsaI-n). The results revealed that the crossbred cows belonged to the RsaI patterns namely b, k, l, a/l, d/s, l/n, l/o and m/n, whose corresponding frequencies were 0.027, 0.013, 0.040, 0.027, 0.040, 0.067, 0.027 and 0.067, respectively. Digestion of the 284 bp PCR product of DRB3.2 gene with PstI in the crossbred cattle did not reveal any restriction site. These results suggested the absence of the recognition site in some of the animals. These results also revealed that the crossbred cows studied were in homozygous as well as heterozygous condition. On the basis of the above results it can be concluded that the DRB3.2 gene was found to be highly polymorphic in the crossbred cattle population.

Effect of Alpha-lactalbumin Gene Polymorphism on Milk Production Traits in Water Buffalo

Dayal, S.,Bhattacharya, T.K.,Vohra, V.,Kumar, P.,Sharma, Arjava Asian Australasian Association of Animal Productio 2006 Animal Bioscience Vol.19 No.3

A genetic study was conducted to elucidate the effect of alpha-Lactalbumin (${\alpha}$-LA) gene polymorphism on milk production traits involving total milk yield and daily milk yield during first lactation in two breeds of water buffaloes namely, Murrah and Bhadawari. Single strand conformation polymorphism (SSCP) was carried out to explore genetic polymorphism present at this locus. For this study, exon 1 region of ${\alpha}$-LA was analyzed. Finally, polymorphism data was associated with milk production traits by employing least square analysis. In Murrah buffalo, five genotypes such as AB, BB, BC, CC and CD and four alleles A, B, C and D were detected whereas in Bhadawari buffalo two genotypes namely, AB and BC and three alleles namely, A, B and C were found. Genotypes showed significant effects ($p{\leq}0.05$) on total milk yield and daily milk yield in Bhadawari buffalo but had non-significant effects on these traits in Murrah buffalo.

PCR-SSCP of Serum Lysozyme Gene (Exon-III) in Riverine Buffalo and Its Association with Lysozyme Activity and Somatic Cell Count

Sahoo, Nihar Ranjan,Kumar, Pushpendra,Bhushan, Bharat,Bhattacharya, T.K.,Sharma, Arjava,Dayal, Sanker,Pankaj, Prabhat Kumar,Sahoo, Monalisa Asian Australasian Association of Animal Productio 2010 Animal Bioscience Vol.23 No.8

Serum lysozyme gene is one of the important genes influencing the immune system as its product can cause lysis of bacterial cell wall by cleaving the peptidoglycan layer. The present investigation on the serum lysozyme gene of Indian riverine buffalo was undertaken with the objectives to identify and characterize single nucleotide polymorphic patterns by PCR-SSCP method as well as to study the effect of different genotypes on serum lysozyme activity and somatic cell count. A total of 280 animals comprising four different famous bubaline breeds (Murrah, Mehsana, Surti and Bhadawari), spread over six different farms across the country were used for this study. A 276 bp (partial intron 2, complete exon 3 and partial intron 3) fragment of lysozyme gene was screened for polymorphism using the SSCP technique. Four genotypes namely AA, AB, BC and AC were observed, out of which BC genotype was found to be the most frequent. Among these three alleles, C allele (0.38) was most prevalent in these populations. Various SSCP allelic variants were cloned for sequencing and sequences were submitted to NCBI Genbank. From the alignment of the nucleotide sequences of various allelic variants, it was found that there were differences in 12 positions among the alleles, out of which maximum variation (at 8 places) was found in the intronic region. The allele A was closer to allele-C than allele-B. Allele B was phylogenetically equidistant from both of the other alleles. Mean lysozyme activity determined in serum samples of different animals of Murrah buffalo was $27.35{\pm}2.42\;{\mu}g$ per ml of serum, whereas the mean somatic cell count was $1.25{\pm}0.13{\times}10^5$ cells per ml of milk. The SSCP pattern-wise effects of various genotypes on lysozyme activity and SCC were analyzed. Although the mean values were apparently different in various genotypes, these differences were statistically non-significant. It can be concluded that the riverine buffaloes are sufficiently polymorphic with respect to serum lysozyme gene. The absence of AA genotype in Bhadawari breed of buffalo can be considered as a marker for breed characterization. The difference of four nucleotides in exon-3 indicates high selection pressure on the gene.

Molecular Characterization of Rathi and Tharparkar Indigenous Cattle (Bos indicus) Breeds by RAPD-PCR

Sharma, Amit Kumar,Bhushan, Bharat,Kumar, Sanjeev,Kumar, Pushpendra,Sharma, Arjava,Kumar, Satish Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.9

Random amplification of polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) analysis was carried out using DNA samples of 30 animals of Rathi cattle and 42 animals of Tharparkar cattle. Genomic DNA was isolated as per standard protocol and evaluated for its quality, purity and concentration. Twenty three random primers were screened out of which 15 primers yielded satisfactory amplifications and were used for further analysis. Average numbers of polymorphic fragments per primer were 7.07${\pm}$0.86 in Rathi and 6.80${\pm}$0.61 in Tharparkar cattle. The percentage of polymorphic bands in these two cattle breeds were 86 and 87%, respectively. Within breed genetic similarities for pooled over primers in the animals of Rathi and Tharparkar breeds were .577${\pm}$0.30 and 0.531${\pm}$0.02, respectively on the basis of band frequency (BF) and 0.645${\pm}$0.04 and 0.534${\pm}$0.04, respectively on the basis of band sharing (BS). Averages of between breed genetic similarities for pooled over primers were 0.97 and 0.92 according to BF and BS, respectively, which reflect higher degree of genetic similarity between Rathi and Tharparkar cattle breeds. Index of genetic distance based on BF and BS for pooled over primers was 0.030${\pm}$0.011 and 0.088${\pm}$0.031, respectively. Percentage of polymorphic bands and within-breed genetic similarities on the basis of band frequency (BF) and band sharing (BS) for pooled over primers revealed higher genetic similarity in Rathi than Tharparkar cattle population. High estimates of between breed genetic similarities for pooled over primers indicated that either Rathi is having decent from Tharparkar or both the cattle breeds are having common descent. Low value of Index of genetic distances between these two cattle breeds may be due to the fact that Rathi and Tharparkar cattle breeds are the native of Thar Desert in Northwest India. The results of between breed genetic distances also confirm the existence of high degree of genetic similarity between these two breeds of cattle.

Effect of Leptin and IGFBP-3 Gene Polymorphisms on Serum IgG Level of Cattle Calves

Choudhary, Vivek,Kumar, Pushpendra,Saxena, V.K.,Bhattacharya, T.K.,Bhushan, Bharat,Sharma, Arjava,Ahmed, K.A. Asian Australasian Association of Animal Productio 2006 Animal Bioscience Vol.19 No.8

Leptin and IGFBP-3 are two proteins that play an important role in growth and metabolism of the animals. They are also involved in the immune function of animals and, thus, are candidate genes for the study of association with immune functions. Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) of these two genes was done to screen 64 crossbred (Holstein Friesian${\times}$Hariana) female calves of one year of age. From each RFLPs (fragments) three genotypes were observed. In all the RFLPs the mutant homozygotes were very less in numbers and, hence, were excluded from the least squares analysis. The serum IgG level was estimated using SRID assay. The mean level of serum IgG was $28.83{\pm}2.73mg/ml$. The effect of these identified genotypes on serum IgG level of calves at one year of age was analysed using least squares analysis. The HaeIII RFLP-AB genotype had significantly (p<0.05) higher serum IgG level ($31.86{\pm}3.05$) than the HaeIII RFLP-AA ($25.62{\pm}2.96$) genotype. There was no significant effect of leptin genotypes on the IgG level. The present results indicated a role of the IGFBP-3 gene on serum IgG level of cattle calves.

Effect of Butyrophilin Gene Polymorphism on Milk Quality Traits in Crossbred Cattle

Bhattacharya, T.K.,Misra, S.S.,Sheikh, Feroz D.,Sukla, Soumi,Kumar, Pushpendra,Sharma, Arjava Asian Australasian Association of Animal Productio 2006 Animal Bioscience Vol.19 No.7

A genetic polymorphism study on butyrophilin gene was carried out to explore variability of this gene and to estimate effects of such variability on milk quality traits in crossbred cattle. Polymorphism was unraveled by conducting Hae III PCR-RFLP of this gene. Three genotypes such as AA, BB and AB and two alleles namely A and B were observed in crossbred population. The frequencies of genotypes and alleles were 0.78, 0.17 and 0.04 for AA, AB and BB genotypes, respectively, and 0.87 and 0.13 for A and B alleles, respectively. The nucleotides, which have been substituted from allele A to B, were observed as C to G ($71^{st}$ nucleotide), C to T ($86^{th}$ nucleotide), A to T ($217^{th}$ nucleotide), G to A ($258^{th}$ nucleotide), A to C ($371^{st}$ nucleotide) and C to T ($377^{th}$ nucleotide). The nucleotide substitutions at $71^{st}$, $86^{th}$ and $377^{th}$ position of the fragment were found as silent mutations whereas nucleotide changes at $217^{th}$, $258^{th}$ and $371^{st}$ positions were detected as substitution of amino acid lysine with arginine, valine with isoleucine, and leucine with proline from allele A to B. The genotypes had significant effects ($p{\leq}0.05$) on total milk solid%, fat%, SNF%, while showing nonsignificant impact on total protein%. AA genotype produced highest average yield for all the traits.

Growth Hormone Gene Polymorphism and Its Effect on Birth Weight in Cattle and Buffalo

Biswas, T.K.,Bhattacharya, T.K.,Narayan, A.D.,Badola, S.,Kumar, Pushpendra,Sharma, Arjava Asian Australasian Association of Animal Productio 2003 Animal Bioscience Vol.16 No.4

The study was carried out in Sahiwal, Holstein Friesian, Jersey and crossbred cattle and Murrah, Bhadwari, Jaffarabadi, Nagpuri and Surti buffaloes maintained at different organized herds to work out the polymorphism at growth hormone locus and study its effect on birth weight. A 223 bp fragment of the gene was amplified and digested with Alu I restriction enzyme. Two alleles, L and V with three genotypes LL, LV and VV were observed in Jersey, Holstein and cross bred cattle. Sahiwal cattle and buffalo were monomorphic for this locus producing only one genotype LL and one allele L. The frequency of L allele was comparatively higher in Holstein and crossbred cattle while in Jersey breed, the frequency of this allele was intermediate. The effect of genotype on birth weight was significant and LV genotype had higher birth weight than other genotypes. Hence, LV genotype in Holstein Friesian favored higher birth weight.