Total RNA was extracted from cattle testes and reverse transcribed and amplified by RT2PCR. The two splice variants of LDH2Cwere obtained. Compared with the full length ORFof bovine LDH2C,as a result,The two cloned splice variants were missing exons,In detail,variants 1(missing the coding sequence of exon 7);Variant 2(missing the coding sequence of exons 4 and 7)lacks a large part of the NAD binding domain and the entire active site. Native poly2 acrylamide gel electrophoresis was performed to determine if the splice variants can be translated into proteins. However, native PAGE detected no specific bands from cattle testes and spermatozoa. This study suggests the alternative splicing of LDH2C might be involved in regulation of LDH2C expression.
HoNG Jian,, ZHENG Yu cai, HE Qing hua
. Cloning and Sequence Analysis of Multiple Splice Variants of Lactate Dehydrogenase C in Cattle Testes[J]. Acta Agriculturae Boreali-Sinica, 2009
, 24(3)
: 51
-53
.
DOI: 10.7668/hbnxb.2009.03.011
[1] Markert C L,Shaldee J B,Whitt G S.Evolution of a gene.Multiple genes for LDH isozymes provide a model of the evo-lution of gene structure,function and regulation[J].Science,1975,189(4197):102-114.
[2] Coonrod S,Vitale A,Duan C,et al.Testis-specific lactate de-hydrogenase(LDH-C4;Ldh3)in murine oocytes and preim-plantation embryos[J].J Andrel,2006,27(4):502-509.
[3] Tsuji S,Qureshi M A,Hou E W,et al.Evolutionary relation-ships of lactate dehydrogenases(LDHs)from mammals,birds,an amphibian,fish,barley,and bacteria:LDH cDNA se-quences from Xenopus,pig,and rat[J].Proc Nail Acad Sci,1994,91(20):9392-9396.
[4] Blanco A,Burgos C,Gerez D E,et al.Properties ofthe testic-ular lactate dehydregenase isoenzyme[J].J Biol Chem,1976,153(2):165-172.
[5] 洪键,贺庆华,郑玉才,等.睾丸特异乳酸脱氢酶-C基因存在选择性剪接[J].自然科学进展,2008,18(9):1070-1074.
[6] Dietz A A,Lubrano T.Separation and quantitation of lactic dehydrogenase isoenzymes by disc electrophoresis[J].Anal Bioehem,1967,20:246-257.
[7] Koslowski M,Tureci O,Bell C,et al.Multiple splice variants of lactate dehydrogenase C selectively expressed in human cancer[J].Cancer Res,2002,62(22):6750-6755.
[8] Kroft T L,Jethanandani P,McLean D J.et al.Methylation of CpG dinucleotides alters binding and silences testis-specific transcription directed by the mouse lactate dehydrogenase C promoter[J].Biol Reprod,2001,65(5):1522-1527.
[9] Jethanandani P,Goldberg E.Ldhc expresdon in non-germ cell nuclei is repressed by NF-1 binding[J].J Biol Chem,2001,276(38):35414-35421.
