Construction of Controllable Expression Vector of Mus musculus Growth Hormone (GH) shRNA and Its Gene Si- lencing Efficiency Detection

Growth hormone （GH） is a polypeptide hormone secreted by the pituitary, which promotes animal growth, muscle development, metabolism regulation and other important physiological functions. In this study, a pair of mGH short hairpin RNA （shRNA） was designed according to mouse （ Mus musculus） GH mRNA sequence; pSingle-tTS-mGH shRNA-RFP, an integrated controllable expression vector of mGH shRNA, was constructed successfully. The recombinant vector was transfected into mouse pituitary tumor cell line AtT-20. After addition of doxycyelin （ DOX）, the expression of red fluorescent protein （RFP） was observed un- der a fluorescent microscope. The expression level of mGH in cells was detected by quantitative Realtime PCR （qRT-PCR） and Western blot. After DOX induction, the relative expression level of GH mRNA in cells transfected with GH shRNA was reduced by about 70% compared with that in DOX-free group and other control groups, exhibiting extremely significant differences （P 〈 0.01 ） ; moreover, the relative expression level of GH protein was reduced by about 90% ; the expression level of GH mRNA and GH protein exhibited no significant difference among other groups （P 〉 0.05）. In this study, a controllable expression vector of GH shRNA with high gene silencing efficiency was constructed successfully, which could be used to reveal GH autocfine / paracrine interactions and analyze functions of GH gene in growth, development and disease occurrence of animals by regulating GH expression levels.

Dietary Control of Lactase Expression in the Weaning Rat

The decline in lactase activity during weaning has been well established. However, its molecuIar mechanism remains to be explored. We studied changes in the expression of lactase in terms of the transcription and translation processes in rat microvillus membrane by Northern blot and Western blot analysis, respectively. To examine the effect of dietary change from a milk to a non-milk diet on the developmental pattern of lactase expression, weaning was prevented by keeping the rats under suckling conditions for 27 days after birth. This treatment only suppressed the extent of decline: while the weanlings showed 17 percent activity compared to that of 4-day-old rats, the prolonged suckling rats showed only 42 percent. The changes in the expression of lactase mRNA and protein were parallel with the change of lactase activity. In other words, the fundamental pattern of significant depression of lactase expression occurred relatively independent of dietary modification.This observation indicates that the regulation of lactase expression is firmly determined at the transcriptional level, and that dietary factor such as the termination of lactose ingestion has only a relatively minor effect

How to Analyze the Function of Control Genes in Batches

Functional analysis of control genes is playing an important role in animal development.The research is mainly focused on developing a method of estimating the gene's effect or function in the development of animals.In this article,the gene's expression and control are described by models of quantitative genetics,which include the genetic models about the genes with no interaction and interaction effect.On this basis,a method analyzing the functions of control genes in batches is advanced,the method includes three steps as follows: Firstly,describe the gene's expression and control with multiple regression models in statistic method in different conditions.Secondly,collect the material of gene's polymorphism related to the gene's expression and control.Because,gene's polymorphism or the codominant molecular marker electropherogram in a gene locus present 3 states,and can be expressed with -1,0 and 1 respectively.The author thinks it can be regarded as levels in orthogonal layout.Discard the materials of gene's polymorphism not fitting the orthogonal layout,and use the materials of gene's polymorphism fitting the orthogonal layout as the data to estimate the gene's effects.Thirdly,analyze the gene's effect or function with equations as follows:suppose that a quantitative trait is controlled by N gene loci,in each locus there are two alleles,the alleles' effects are A GM1 ?a GM1 ;A GM2 ? a GM2 ;...;A GMi ?a GMi ;...;A GMN ;a GMN units respectively.When the genotype in a locus is heterozygous A GMi a GMi ,the interaction coefficient of alleles in a locus is K units.If there is no interaction among different gene loci,the author thinks,the gene's effects and gene's type can be estimated by using such equation: P GM =2∑Ni=1(A i-a i+K i)+∑Ni=1(A i-a i)M GMi -∑Ni=1K GMi M GMi 2 If there are interactions among different gene loci,and the effect of the x th gene's state combined with the y th gene's state in the k th gene group is defined as Y xyr ,and let gene group electropherogram states be r (where r∈ ),the author thinks,the gene group's effects can be estimated by the equation: Y xyr = K PCxy0 +K PCxy r+K PCxy' r 2+K PCxy'' 3+K PCxy(3') r 4+K PCxy(4') r 5+K PCxy(5') r 6+K PCxy(6') r 7+K PCxy(7') r 8 If there are interactions among more genes,the analogous method can also be used. In this way,be using the gene's codominent marker materials fitting the orthogonal layout,the gene's expression and control can be researched by the method of multiple regression,the gene's relative effect can be estimated,the overdominant gene's overdominant coefficient can be assessed,the gene's type can be identified,the interaction gene group's relative effect and the interaction gene's relation can be estimated as well,and furthermore,the gene's relation can be expressed in equations.For the sake of understanding the method used in this paper,the rout to prove it is given as well.Finally,the author thinks,this method is very useful to research the gene's function in molecular developmental biology.

JNK1 negatively controls antifungal innate immunity by suppressing CD23 expression

Subject Code:H10 With the support by the National Natural Science Foundation of China,a study by Dr.Lin Xin(林欣)’s groups from the Institute for Immunology,School of Medicine,Tsinghua University demonstrates that JNK1negatively regulates CD23,a novel C-type lectin receptor(CLR)with antifungal properties。